- development of antiviral peptides,
- modification of antibody flexible chane
to virus (see previus research)
- stepwise testing of each antibody to antigen,
- determination of key amino acid residue
- modification of antibody flexible chane
to virus (see previus research)
- stepwise testing of each antibody to antigen,
- determination of key amino acid residue
Our method include:
- range of affinity changes,
- determining the effectiveness of drugs due to mutations in virus (see previus research)
- definition of resistance,
- range of affinity changes,
- determining the effectiveness of drugs due to mutations in virus (see previus research)
- definition of resistance,
AI
Development of virus inhibitors taking into account the mutational variability of viruses
We are open for collaboration for the implementation of innovative technology.
The goal of our work is to create a method that would allow experimenters to obtain visual physical characteristics of biochemical reactions in an understandable accessible format.
At the same time, we set ourselves the goal that the obtained data could be directly compared with the data obtained in an in vitro experiment.
Modeling the impact of point mutations on the stability of proteins. EXAMPLE 1 BIOLOGICAL SOFT
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Results of performed studies on finding a peptide inhibitor to the Spike protein
the value lg(cond(W)) characterizes the stability of the formed biological complex.
This value is directly proportional to Kd, the larger it is, the lower the affinity.
This value is directly proportional to Kd, the larger it is, the lower the affinity.
We are interested in the smallest value of this value in relation to the value of the interaction of wild-type proteins.
Its interaction N2 with substitutions
997MET1002MET.
Its interaction N2 with substitutions
997MET1002MET.
the value delta(H) indicates a change in the degree of ordering of the system during the transition from one state to the next state.
we are interested in the negative value of entropy in the negative range.
The graph shows that this condition corresponds to the value number 2.
we are interested in the negative value of entropy in the negative range.
The graph shows that this condition corresponds to the value number 2.
The information on the page below provides a more detailed breakdown of the study with graphs and charts.
Amino acid residue substitutions are made in the short peptide to improve binding to the spike protein of the coronavirus.
The contribution of
point mutations to the affinity of a dimeric protein complex
point mutations to the affinity of a dimeric protein complex
ACE2- (S) -glycoprotein dimeric complex was investigated and also research the identification of key amino acid residues in the ACE2 protein when binding to the SARS-CoV-2 virus
Bellow we present the results of mathematical simulation of the the contribution of point mutations to the affinity of a dimeric protein complex. As an example, the ACE2- (S) -glycoprotein dimeric complex was investigated and research the identification of key amino acid residues in the ACE2 protein when binding to the SARS-CoV-2 virus Spike-glycoprotein viral protein; the replacement of amino acid residues in the ACE2 protein in the alfa-helical region on the N-end side that participate in binding to the (S) glycoprotein was performed.
The method developed allows us to analyze the change in the affinity of a dimeric protein complex during mutations in the region of the active center in any of the proteins. The results obtained are in good agreement with the methods in [Stawiski E.W., Diwanji D. et al. Human ACE2 receptor polymorphisms predict SARS-CoV-2 susceptibility //BioRxiv 2020, doi: https://doi.org/10.1101/2020.04.07.024752.]
The method developed allows us to analyze the change in the affinity of a dimeric protein complex during mutations in the region of the active center in any of the proteins. The results obtained are in good agreement with the methods in [Stawiski E.W., Diwanji D. et al. Human ACE2 receptor polymorphisms predict SARS-CoV-2 susceptibility //BioRxiv 2020, doi: https://doi.org/10.1101/2020.04.07.024752.]
This article describe a detailed method for selecting inhibitors based on modified natural peptides for the SARS-CoV protein spike glycoprotein. The selection of inhibitors is carried out by increasing the affinity of the peptide to the active center of the protein. The article also provides a step-by-step guide to the analysis of affinity of interaction by comparing 3 criteria, presents an analysis of energy interactions between the active center of a protein and the wild-type peptide interacting with it and taking into account modifications of the latter.
Fig.1.Interactions between SARS-CoV-2- and ACE2. The ACE2 mainly engages the alfa-helix in the recognition of the spike-glycoprotein.
Fig. 1. shows the site of interaction of the two proteins SARS-CoV-2 and ACE2, which we will study, namely, the alfa-spiral site from 24 a.a. to 38 a.a.
We simulated the substitution of amino acid residues in the wild type Ace2 protein in the alfa-helix region and analyzed the influence of such substitutions on the change in stability of the protein complex Ace2-(S)-glycoprotein.
In the process of analyzing changes in the interaction of two proteins during dimer formation, we will analyze two values that characterize the stability of the protein complex
1.The condition number condition number lg(cond(W)) of the matrix the elements of which are potential energies of electrostatic interaction between pairs of amino acid residues in proteins.
2. For a more accurate estimate characterizing the stable state of the biological complex, it is necessary to correlate the parameter lg(cond(W))with the change in the differential
entropy measure (delta)H.
Moreover, if the first quantity lg(cond(W)) is characterized by the minimum value, and the second (delta) H by a negative value, and the observed dynamics of change (transformation) of these two quantities occur simultaneously (synchronously), then this will indicate a transition of the biological system to a more stable state.
We simulated the substitution of amino acid residues in the wild type Ace2 protein in the alfa-helix region and analyzed the influence of such substitutions on the change in stability of the protein complex Ace2-(S)-glycoprotein.
In the process of analyzing changes in the interaction of two proteins during dimer formation, we will analyze two values that characterize the stability of the protein complex
1.The condition number condition number lg(cond(W)) of the matrix the elements of which are potential energies of electrostatic interaction between pairs of amino acid residues in proteins.
2. For a more accurate estimate characterizing the stable state of the biological complex, it is necessary to correlate the parameter lg(cond(W))with the change in the differential
entropy measure (delta)H.
Moreover, if the first quantity lg(cond(W)) is characterized by the minimum value, and the second (delta) H by a negative value, and the observed dynamics of change (transformation) of these two quantities occur simultaneously (synchronously), then this will indicate a transition of the biological system to a more stable state.
Let us consider in more detail the obtained results for peptide inhibitors on viral activity.
1. Previously reported SARS-CoV infectivity was reduced over 10,000-fold through pre-incubation with two peptides, while it was completely inhibited in the presence of three peptides.Were reported that four 20-mer synthetic peptides (S protein fragments), designed to span these sequence variation hotspots, exhibited significant antiviral activities in a cell line. [Synthetic peptides outside the spike protein heptad repeat regions as potent inhibitors of SARS-associated coronavirus]
2. Another study reported that Peptide P8 (P2 + P6 + P10) exhibited the strongest antiviral activity, Synergistic antiviral effects mediated by peptide combinations. [Synthetic peptides outside the spike protein heptad repeat regions as potent inhibitors of SARS-associated coronavirus]
3. In the study [Synthesized peptide inhibitors of HIV-1 gp41-dependent membrane fusion. Curr Pharm Des 390 19: 1800-9] peptides derived from the HR1 and HR2 sequences of the class I viral fusion proteins have been demonstrated to possess antiviral activity through binding to the prehairpin intermediate thus blocking the formation of viral 6-HB core.
4. Cholesterylated peptide exhibits greatly increased α-helical stability and target-binding affinity [Design of 1 potent membrane fusion inhibitors against SARS-CoV-2, an emerging coronavirus with high fusogenic activity]
5. Successful peptide inhibitors of virus penetration into the host cell have been successfully developed in the case of HIV-1, Ebola virus, paramyxoviruses (SV5), respiratory syncytial virus (RSV). So, the Ebola virus was 99% inhibited when using the HIV peptide C-peptide conjugate as an inhibitor. The sequence found is characterized by the highest activity against the Ebola virus [Inhibition of Ebola Virus Entry by a C-peptide Targeted to Endosomes]
6. Inhibitor molecules included D peptides, synthetic peptides from the N and C ends of a protein virus molecule that could bind the HR1 viral domain and effectively inhibit viral infection.
In this paper, we propose a stepwise increase in the affinity of a natural peptide for a target viral protein. we have developed a phased, five-point method for determining the most suitable modified peptide. The method also includes a phased selection of the alpha-helical region for such modifications that lead to an increase in the stability of the dimer complex compared to the wild-type protein dimer, selection criteria are presented and described.
2. Another study reported that Peptide P8 (P2 + P6 + P10) exhibited the strongest antiviral activity, Synergistic antiviral effects mediated by peptide combinations. [Synthetic peptides outside the spike protein heptad repeat regions as potent inhibitors of SARS-associated coronavirus]
3. In the study [Synthesized peptide inhibitors of HIV-1 gp41-dependent membrane fusion. Curr Pharm Des 390 19: 1800-9] peptides derived from the HR1 and HR2 sequences of the class I viral fusion proteins have been demonstrated to possess antiviral activity through binding to the prehairpin intermediate thus blocking the formation of viral 6-HB core.
4. Cholesterylated peptide exhibits greatly increased α-helical stability and target-binding affinity [Design of 1 potent membrane fusion inhibitors against SARS-CoV-2, an emerging coronavirus with high fusogenic activity]
5. Successful peptide inhibitors of virus penetration into the host cell have been successfully developed in the case of HIV-1, Ebola virus, paramyxoviruses (SV5), respiratory syncytial virus (RSV). So, the Ebola virus was 99% inhibited when using the HIV peptide C-peptide conjugate as an inhibitor. The sequence found is characterized by the highest activity against the Ebola virus [Inhibition of Ebola Virus Entry by a C-peptide Targeted to Endosomes]
6. Inhibitor molecules included D peptides, synthetic peptides from the N and C ends of a protein virus molecule that could bind the HR1 viral domain and effectively inhibit viral infection.
In this paper, we propose a stepwise increase in the affinity of a natural peptide for a target viral protein. we have developed a phased, five-point method for determining the most suitable modified peptide. The method also includes a phased selection of the alpha-helical region for such modifications that lead to an increase in the stability of the dimer complex compared to the wild-type protein dimer, selection criteria are presented and described.
The structure of the Spike-glycoprotein
Three-dimensional structure of the dimeric complex of the peptide "blue" Spike-glycoprotein and the "green" protein Spike-glycoprotein. We will replace amino acid residues in the "blue" peptide. Interacting sites are represented by alpha-helical sites.
Fig.2. Three-dimensional structure of the dimeric complex of the peptide "blue" Spike-glycoprotein and the "green" protein Spike-glycoprotein. We will replace amino acid residues in the "blue" peptide. Interacting sites are represented by alpha-helical sites.
Method of selection of inhibitors to the active center of the Spike-glycoprotein protein.
In this section, we will describe in detail the step-by-step instructions for determining key amino acid residues in the interaction of two proteins, methods for increasing the affinity of a protein dimer by modifying the polypeptide chain of one of the proteins, see Fig. 3 (schemas)
To maximize compliance with the secondary structure, we will investigate alpha-helical regions. The analyzed quantities in this case are log (cond (w)) and ∆H.
To maximize compliance with the secondary structure, we will investigate alpha-helical regions. The analyzed quantities in this case are log (cond (w)) and ∆H.
Fig.3. Scheme of stepwise selection and modification of a natural peptide to increase the affinity of the dimeric complex
- №Step1DescriptionSelection of three-dimensional structures of target proteins (dimers / tetramers). We focused on the data obtained using X-ray diffraction analysis of the Spike-glicoproteins trimeric complex (PDB: 5x5b). We opted for alpha-helical structures, which may contain interacting amino acid residues of two proteins.
- №Step2DescriptionControl and verification of the received data can be carried out in two ways:
-to perform a calculation for two different three-dimensional structures of the same biological complex,
- perform a random experimental check, or use the previously available experimental data on the effect of substitutions of amino acid residues on affinity of binding - №Step3DescriptionMutagenesis of amino acid residues of one of the alpha-helical structures of the Spike-glycoprotein protein
3.1. Calculation of the potential energy of pairwise interaction of all amino acid residues of one protein with amino acid residues of another protein for wild-type proteins. The formation of the obtained values in the form of a matrix.
3.2. Calculation of the potential energy of pairwise interaction of the dimeric protein complex, taking into account each replacement of the amino acid residue in one of the participant's polypeptide chains. The formation of the obtained energy values in the form of a matrix.
3.3. Determination of lg (cond (W)) for the interaction matrix of wild-type proteins and mutant forms of proteins.
3.4. Determination of ΔH during the transition of the system and from the interaction of mutant forms of the protein to the interaction of wild-type proteins.
3.5. Comparison of the obtained data log (cond (W)) and ∆Н in the form of graphs.
3.6. Analysis of the obtained data: the smallest value of lg (cond (W)) and the corresponding negative ∆Н. - №Step4DescriptionThe determination of key amino acid residues, the replacement of which leads to an increase in the stability of the dimeric complex, often, these amino acid residues correspond to high potential energies of pairwise electrostatic interaction between the amino acid residues of the two proteins. We think that the surrounding values of the interaction energies of neighboring residues play an important role here. We get a set of key amino acid residues, according to the implementation of paragraph 3.6.
- №Step5DescriptionWe replace several identified amino acid residues at the same time, compare the minimum values of lg (cond (W)) and the negative values of ∆H, repeat all the steps from Step 3, but only for several substitutions at the same time: finding the potential energy of pairwise interaction, forming a matrix, calculating lg ( cond (w)) and ΔН
At the end of the calculations, we select those replacements / modifications that meet two verification criteria: the minimum values of lg (cond (W)) and the maximum negative values of ∆H. We will talk about the third criterion in more detail in the discussion section of the results.
Fig.4. A clear diagram of the analysis of changes in the potential interaction energy when replacing C1014M. Amino acid sequence of the "blue" Spike-glycoprotein a), three-dimensional structure of two interacting alpha-helical sections of the dimer b), graph of fluctuations in the potential energy of pairwise electrostatic interaction when replacing C1014M c), three-dimensional representation of the potential energy of electrostatic pairwise interaction of two truncated wild-type proteins with indication of some numerical values of the interaction d), three-dimensional representation of the potential energy of electrostatic pairwise interaction of two truncated proteins, the C1014M was replaced in the "blue" peptide, some numerical values are indicated by arrows energy e)
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, or 2019-nCoV) is an enveloped non-segmented positive-sense RNA virus
Two quantities lg (cond (W)) and ∆H help us estimate this change. Moreover, if the value of lg (cond (W)) decreases after replacing the amino acid residue, this may signal an increase in the stability of the dimer complex.
We replace C1014M in the "blue" Spike-glycoprotein peptide when interacting with the "yellow" Spike-glycoprotein, the results are shown in Fig. 4. When replacing hydrophilic cysteine with hydrophobic methionine, we see significant changes in the potential energy values, in particular, a decrease in the interaction energy of methionine with other amino acid residues of the second protein.
Fig. 6c shows a comparative graph of the change in the potential energy of interaction between C1014 and M1014 with other amino acid residues of the second protein.
The significance of such change in the potential interaction energy when replacing cysteine with methionine must be analyzed, taking into account the values of the energy interactions of neighboring amino acid residues.
Two quantities lg (cond (W)) and ∆H help us estimate this change.
Fig. 6c shows a comparative graph of the change in the potential energy of interaction between C1014 and M1014 with other amino acid residues of the second protein.
The significance of such change in the potential interaction energy when replacing cysteine with methionine must be analyzed, taking into account the values of the energy interactions of neighboring amino acid residues.
Two quantities lg (cond (W)) and ∆H help us estimate this change.
Fig.5.Schedule of amino acid residue substitutions in the "blue" Spike-glycoprotein interacting with the Spike-glycoprotein. The calculation was performed for the structure shown in Fig.1
The results of the numerical calculations presented on the graph allow us to conclude that some substitutions of amino acid residues starting from 994 a.a. to 1014 a.a. can significantly affect the change in the affinity of the dimeric complex.
The results of the numerical calculations presented on the graph allow us to conclude that some substitutions of amino acid residues starting from 994 a.a. to 1014 a.a. can significantly affect the change in the affinity of the dimeric complex.
Fig.6. Dependence of lg (cond (W)) on the successive replacement of the amino acid residue in the "blue" peptide with MET (M), CYS (C), PHE (F), VAL (V), SER (S) during the formation of the dimeric complex.
Since we are only interested in those modifications that can increase the stability of the dimeric complex, we dwell in more detail on the selection criteria:
1. The minimum value of lg (cond (W))
2. The corresponding negative value of the measure of change in differential entropy ∆Н
Since we are only interested in those modifications that can increase the stability of the dimeric complex, we dwell in more detail on the selection criteria:
1. The minimum value of lg (cond (W))
2. The corresponding negative value of the measure of change in differential entropy ∆Н
| Fig.7. The results of numerical calculations of changes in the differential entropy measure during the transition of a system from a modified state to a state of interaction of wild-type proteins. In the corner of each graph, the number of the replaced amino acid residue in the blue Spike-glycoprotein peptide is additionally indicated. Each of the presented amino acid residues was alternately replaced by MET (M), CYS(C), VAL (V), SER (S). |
Fig.8. The results of numerical calculatios of substitutions of amino acid residues from 1011a.a. to 1014 a.a. alternately with MET (M), CYS (C), PHE (F), VAL (V), Ser (S).
As can be seen from the graphs in Fig. , each replacement of the amino acid residue with cysteine leads to a decrease in the stability of the dimeric complex and to an increase in the measure of change in differential entropy. Thus, we do not recommend the use of cysteine substitutions of these amino acid residues to increase the affinity of the dimeric complex.
As can be seen from the graphs in Fig. , each replacement of the amino acid residue with cysteine leads to a decrease in the stability of the dimeric complex and to an increase in the measure of change in differential entropy. Thus, we do not recommend the use of cysteine substitutions of these amino acid residues to increase the affinity of the dimeric complex.
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Population-based serosurveys measuring anti-severe acute respiratory syndrome coronavirus 2 (anti-SARS-CoV-2) antibodies provide. IgM and IgG antibodies to SARS-CoV-2 are generally detectable in blood several days after initial infection, although the duration of time antibodies are present. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, or 2019-nCoV) is an enveloped non-segmented positive-sense RNA virus/ Qualitative detection of IgG antibodies to SARS-CoV-2, the virus that causes COVID-19, to help identify individuals
Fig.9. The results of numerical calculations of the change in the measure of differential entropy (delta)H when alternately replacing the amino acid residues from 1011 a.a. to 1014 a.a. with MET (M), CYS (C), PHE (F), VAL (V), SER (S).
The most significant result was obtained with the replacement of 1014 a.a. on MET (M), PHE (F), VAL (V), SER (S), since all the obtained ∆H values are in the negative range of values, the value log (cond (W)) is also characterized by values in a lower range of values .
The most significant result was obtained with the replacement of 1014 a.a. on MET (M), PHE (F), VAL (V), SER (S), since all the obtained ∆H values are in the negative range of values, the value log (cond (W)) is also characterized by values in a lower range of values .
Population-based serosurveys measuring anti-severe acute respiratory syndrome coronavirus 2 (anti-SARS-CoV-2) antibodies provide. IgM and IgG antibodies to SARS-CoV-2 are generally detectable in blood several days after initial infection, although the duration of time antibodies are present. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, or 2019-nCoV) is an enveloped non-segmented positive-sense RNA virus/ Qualitative detection of IgG antibodies to SARS-CoV-2, the virus that causes COVID-19, to help identify individuals
The fifth final stage
At the fifth final stage, we have to select the amino acid residues in the wild-type peptide of the blue Spike-glycoprotein, which we will modify. In this work, the primary task was to disclose the methodology for the stepwise selection and modification of peptides to increase affinity for the second protein.
The finding of a high affinity peptide is shown here as an example on several amino acid residues. In order to find all possible modifications of the peptide, it will be necessary to carry out a similarly extensive numerical study, which is beyond the scope of this work.
We took 4 amino acid residues: 997, 998, 1002 and 1006, made replacements for them with Met, GLN, ASN, obtained potential energy matrices, calculated the values of Lg (cond (W)) and ∆H. The results of the calculations are shown in Fig. 21
The finding of a high affinity peptide is shown here as an example on several amino acid residues. In order to find all possible modifications of the peptide, it will be necessary to carry out a similarly extensive numerical study, which is beyond the scope of this work.
We took 4 amino acid residues: 997, 998, 1002 and 1006, made replacements for them with Met, GLN, ASN, obtained potential energy matrices, calculated the values of Lg (cond (W)) and ∆H. The results of the calculations are shown in Fig. 21
Population-based serosurveys measuring anti-severe acute respiratory syndrome coronavirus 2 (anti-SARS-CoV-2) antibodies provide. IgM and IgG antibodies to SARS-CoV-2 are generally detectable in blood several days after initial infection, although the duration of time antibodies are present. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, or 2019-nCoV) is an enveloped non-segmented positive-sense RNA virus/ Qualitative detection of IgG antibodies to SARS-CoV-2, the virus that causes COVID-19, to help identify individuals
Fig.10. A graphical representation of the numerical results of numerical modeling of several simultaneous modifications of amino acid residues in the blue Spike-glycoprotein at 997, 998, 1002, 1006, 1008 amino acid residues. Substitutions were made with MET, GLN, ASN.
Change in lg (cond (W)) a), change in ∆Н b).
Thus, the conclusions drawn from numerical calculations on the modification of the natural peptide allow us to conclude that the simultaneous replacement of amino acid residues 997MET and 1002MET can lead to an increase in the affinity of the dimeric complex formed by the modified natural viral peptide and the Spike-glycoprotein protein. Since the affinity of such a modified complex may exceed the interaction of wild-type viurs proteins, this method will allow finding inhibitors by the competitive type of binding
Change in lg (cond (W)) a), change in ∆Н b).
Thus, the conclusions drawn from numerical calculations on the modification of the natural peptide allow us to conclude that the simultaneous replacement of amino acid residues 997MET and 1002MET can lead to an increase in the affinity of the dimeric complex formed by the modified natural viral peptide and the Spike-glycoprotein protein. Since the affinity of such a modified complex may exceed the interaction of wild-type viurs proteins, this method will allow finding inhibitors by the competitive type of binding
antibodies against SARS-CoV -2, the virus which causes the disease COVID-19 in general, a positive antibody test is presumed to mean a person has been infected with SARS-CoV-2, the virus that causes COVID-19 Roche Diagnostics has developed a serology test for detecting antibodies to the severe acute respiratory syndrome coronavirus 2 that causes COVID-19 Studies from serum samples could transform our understanding of the spread of COVID-19, but what antibodies alone say about immunity Antibody responses against enveloped viruses, such as SARS‐CoV‐2, are usually comprised of immunoglobulin (Ig) M, IgG3, IgG1 and IgA COVID-19 is an infectious disease caused by the SARS-CoV-2 virus that spreads easily between people in a similar way to the common
Discussions and conclusions.
_______This article described a method for phasing the affinity of a natural peptide to a target viral protein by modifying the peptide. The selection criteria for the most suitable modifications were given, among which the following can be listed: the stability of the dimer complex by calculating the value of log (cond (W)) and comparing the obtained value with the value of log (Cond (W)) obtained for the wild-type dimer and other modified dimers .
_______The next indicator was the measure of the change in differential entropy ∆Н, which characterizes the ordering of the system. Moreover, the values of lg (cond (W)) and ∆Н are interrelated: the minimum number lg (cond (W)) must correspond to a negative value of ∆Н, otherwise, the dimeric complex does not satisfy the necessary criteria.
________As an example of the selection of suitable modifications of natural peptides, the Spike-glycoprotein viral protein was selected, which is responsible for the attachment of the virus to the cell and the incorporation of the viral genome into the cell. In this work, we analyzed the Central helix region of this protein, performed mutagenesis, and calculated the stability of the formed dimeric complexes upon modification of the natural peptide in the region of the Central helix region. The calculation results showed that the modification of the site from 995 a.a. till 1014 a.a. most likely to affect the affinity of the modified peptide for the Spike-glycoprotein viral protein. In the course of the numerical simulation, wild-type peptide mutagenesis was performed, the calculation of the values of log (cond (W)) and ΔH, the minimum values of log (cond (W)) were compared with negative values of ΔH, since these values are analyzed in pairs. If the log (cond (W)) values take the minimum values, and the ΔH value is in the range of positive values, then this modified peptide does not pass the test according to the criteria.
________To reduce false results, we remove from further analysis all dimeric complexes that are characterized by a ∆H value in the positive range of values. We also believe that an important aspect affecting the results of the modification of a natural peptide is the environment of the modified portion of the polypeptide chain with other amino acid residues, the location of which in turn depends on the solvent (water, impurities, salts, temperature, pH). in three-dimensional space is characterized by the distribution of potential energy of interaction with other amino acid residues in three-dimensional space. Replacing even one amino acid residue in a polypeptide chain can serve as a substantial redistribution of the potential energy of interaction with all amino acid residues of the adjacent polypeptide chain, which in turn will affect the stability of the dimer complex and the entropy of the dimer complex, and more precisely, entail measures to change the differential entropy of the dimer complex ∆H , which we calculate for each replacement.
______By analyzing the Spike-glycoprotein protein peptide in this way, we determined amino acid residues, the replacement of which can affect the affinity, namely, the increase in affinity due to increased stability of the dimeric complex and ordering of the system of two proteins. The aim of this work was to develop and describe in detail a methodology for increasing the affinity of a natural peptide for a target protein by modifying the peptide, namely, replacing amino acid residues. In the course of the work done, several key amino acid residues found were replaced with 997, 998, 1002, 1006, 1008, see fig. on MET (M), GLN (), ASN ().
________The results presented in fig. and in the table it was possible to determine the most suitable modification of the peptide to increase the affinity of interaction with the Spike-glycoprotein viral protein. These modifications are two substitutions ALA997MET and ALA1002 MET, since two simultaneous substitutions significantly reduced the value of log (cond (W)), and the value of ∆Н was characterized by the lowest negative value. A good result was also obtained with the simultaneous replacement of two amino acid residues in the peptide ALA997MET and ALA1002GLN, as well as ALA997MET and ALA1008MET. The worst result was obtained with the simultaneous replacement of ALA998MET and LEU1006GLN, since the value of log (cond (W)) was obtained in a high range of values, and the value of ΔН was in the range of positive values.
To identify a high affinity modified peptide for the wild-type virus virus Spike-glycoprotein, it is necessary to perform large-scale numerical calculations and, based on the results, obtain the most suitable modifications of the peptide for further refinement by increasing the resistance of peptides against protease cleavage and increasing the circulation time in the body to achieve a significant therapeutic effect .
_______The next indicator was the measure of the change in differential entropy ∆Н, which characterizes the ordering of the system. Moreover, the values of lg (cond (W)) and ∆Н are interrelated: the minimum number lg (cond (W)) must correspond to a negative value of ∆Н, otherwise, the dimeric complex does not satisfy the necessary criteria.
________As an example of the selection of suitable modifications of natural peptides, the Spike-glycoprotein viral protein was selected, which is responsible for the attachment of the virus to the cell and the incorporation of the viral genome into the cell. In this work, we analyzed the Central helix region of this protein, performed mutagenesis, and calculated the stability of the formed dimeric complexes upon modification of the natural peptide in the region of the Central helix region. The calculation results showed that the modification of the site from 995 a.a. till 1014 a.a. most likely to affect the affinity of the modified peptide for the Spike-glycoprotein viral protein. In the course of the numerical simulation, wild-type peptide mutagenesis was performed, the calculation of the values of log (cond (W)) and ΔH, the minimum values of log (cond (W)) were compared with negative values of ΔH, since these values are analyzed in pairs. If the log (cond (W)) values take the minimum values, and the ΔH value is in the range of positive values, then this modified peptide does not pass the test according to the criteria.
________To reduce false results, we remove from further analysis all dimeric complexes that are characterized by a ∆H value in the positive range of values. We also believe that an important aspect affecting the results of the modification of a natural peptide is the environment of the modified portion of the polypeptide chain with other amino acid residues, the location of which in turn depends on the solvent (water, impurities, salts, temperature, pH). in three-dimensional space is characterized by the distribution of potential energy of interaction with other amino acid residues in three-dimensional space. Replacing even one amino acid residue in a polypeptide chain can serve as a substantial redistribution of the potential energy of interaction with all amino acid residues of the adjacent polypeptide chain, which in turn will affect the stability of the dimer complex and the entropy of the dimer complex, and more precisely, entail measures to change the differential entropy of the dimer complex ∆H , which we calculate for each replacement.
______By analyzing the Spike-glycoprotein protein peptide in this way, we determined amino acid residues, the replacement of which can affect the affinity, namely, the increase in affinity due to increased stability of the dimeric complex and ordering of the system of two proteins. The aim of this work was to develop and describe in detail a methodology for increasing the affinity of a natural peptide for a target protein by modifying the peptide, namely, replacing amino acid residues. In the course of the work done, several key amino acid residues found were replaced with 997, 998, 1002, 1006, 1008, see fig. on MET (M), GLN (), ASN ().
________The results presented in fig. and in the table it was possible to determine the most suitable modification of the peptide to increase the affinity of interaction with the Spike-glycoprotein viral protein. These modifications are two substitutions ALA997MET and ALA1002 MET, since two simultaneous substitutions significantly reduced the value of log (cond (W)), and the value of ∆Н was characterized by the lowest negative value. A good result was also obtained with the simultaneous replacement of two amino acid residues in the peptide ALA997MET and ALA1002GLN, as well as ALA997MET and ALA1008MET. The worst result was obtained with the simultaneous replacement of ALA998MET and LEU1006GLN, since the value of log (cond (W)) was obtained in a high range of values, and the value of ΔН was in the range of positive values.
To identify a high affinity modified peptide for the wild-type virus virus Spike-glycoprotein, it is necessary to perform large-scale numerical calculations and, based on the results, obtain the most suitable modifications of the peptide for further refinement by increasing the resistance of peptides against protease cleavage and increasing the circulation time in the body to achieve a significant therapeutic effect .
Advantages of ordering with us
-
Cost savingsOrdering work with us, you save almost 90% on preliminary screening experiments to find the most suitable immunoglobulins -
VisibilityThree-dimensional map of the energy interaction of the amino acid sequences of an antibody with an antigen -
No experiment neededDetermination of the affinity range for various substitutions of amino acid residues, during the formation of an antibody-antigen complex before an in vitro experiment -
Wide coverageYou do not have to be limited to a narrow list of substitutions of amino acid residues; instead, you get a wide range of possible modifications -
Affinity range antibodies-antigenThe results obtained will determine the key amino acid residues that make the greatest changes in the affinity of the dimeric complex. -
EfficiencyWe take care about our clients time. Just contact us — and we will help you with all the questions.
Population-based serosurveys measuring anti-severe acute respiratory syndrome coronavirus 2 (anti-SARS-CoV-2) antibodies provide. IgM and IgG antibodies to SARS-CoV-2 are generally detectable in blood several days after initial infection, although the duration of time antibodies are present. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, or 2019-nCoV) is an enveloped non-segmented positive-sense RNA virus/ Qualitative detection of IgG antibodies to SARS-CoV-2, the virus that causes COVID-19, to help identify individuals
