Peter V Dubovskii | Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry (original) (raw)
Papers by Peter V Dubovskii
Toxins
In aqueous solutions, cobra cytotoxins (CTX), three-finger folded proteins, exhibit conformationa... more In aqueous solutions, cobra cytotoxins (CTX), three-finger folded proteins, exhibit conformational equilibrium between conformers with either cis or trans peptide bonds in the Nterminal loop (loop-I). The equilibrium is shifted to the cis form in toxins with a pair of adjacent Pro residues in this loop. It is known that CTX with a single Pro residue in loop-I and a cis peptide bond do not interact with lipid membranes. Thus, if a cis peptide bond is present in loop-I, as in a Pro-Pro containing CTX, this should weaken its lipid interactions and likely cytotoxic activities. To test this, we have isolated seven CTX from Naja naja and N. haje cobra venoms. Antibacterial and cytotoxic activities of these CTX, as well as their capability to induce calcein leakage from phospholipid liposomes, were evaluated. We have found that CTX with a Pro-Pro peptide bond indeed exhibit attenuated membrane-perturbing activity in model membranes and lower cytotoxic/antibacterial activity compared to their counterparts with a single Pro residue in loop-I.
Biochemistry (Moscow), 2014
Electrical charge and amphiphilicity are fundamental characteristics of biomolecules governing th... more Electrical charge and amphiphilicity are fundamental characteristics of biomolecules governing their activity in vivo. Amphiphilic peptides carrying a multitude of electrical charges are physiologically active due to their ability to interact with receptors and a variety of biomembrane components [1, 2]. In particular, polycationic peptides are considered as perspective molecules for antibacterial and anticancer therapy [3, 4]. Such peptides are widely present in the venoms of insects, arachnids, and snakes [5, 6]. Some of them, e.g. cardiotoxins, feature compact and highly stable spatial structure due to the presence of disulfide bonds [7, 8]. In contrast, linear peptides exhibit high flexibility and are typically able to switch among a number of states [9, 10]. In this work we studied amphiphilic peptides with high net positive charge-linear cationic peptides (LCP). These include latarcins Ltc1K [11] and Ltc2a [12] isolated from the venom of the Middle Asia Lachesana
Protein Expression and Purification, 2017
Cytotoxins or cardiotoxins is a group of polycationic toxins from cobra venom belonging to the &#... more Cytotoxins or cardiotoxins is a group of polycationic toxins from cobra venom belonging to the 'three-finger' protein superfamily (Ly6/uPAR family) which includes small β-structural proteins (60-90 residues) with high disulfide bond content (4-5 disulfides). Due to a high cytotoxic activity for cancer cells, cytotoxins are considered as potential anticancer agents. Development of the high-throughput production methods is required for the prospective applications of cytotoxins. Here, efficient approach for bacterial production of recombinant analogue of cytotoxin I from N. oxiana containing additional N-terminal Met-residue (rCTX1) was developed. rCTX1 was produced in the form of E. coli inclusion bodies. Refolding in optimized conditions provided ∼6 mg of correctly folded protein from 1 L of bacterial culture. Cytotoxicity of rCTX1 for C6 rat glioma cells was found to be similar to the activity of wild type CTX1. The milligram quantities of 13C,15N-labeled rCTX1 were obtained. NMR study confirmed the similarity of the spatial structures of recombinant and wild-type toxins. Additional Met residue does not perturb the overall structure of the three-finger core. The analysis of available data for different Ly6/uPAR proteins of snake and human origin revealed that efficiency of their folding in vitro is correlated with the number of proline residues in the third loop and the surface area of hydrophobic residues buried within the protein interior. The obtained data indicate that hydrophobic core is important for the folding of proteins with high disulfide bond content. Developed expression method opens new possibilities for structure-function studies of CTX1 and other related three-finger proteins.
Toxins, 2022
Cobra cytotoxins (CTs) belong to the three-fingered protein family and possess membrane activity.... more Cobra cytotoxins (CTs) belong to the three-fingered protein family and possess membrane activity. Here, we studied cytotoxin 13 from Naja naja cobra venom (CT13Nn). For the first time, a spatial model of CT13Nn with both “water” and “membrane” conformations of the central loop (loop-2) were determined by X-ray crystallography. The “water” conformation of the loop was frequently observed. It was similar to the structure of loop-2 of numerous CTs, determined by either NMR spectroscopy in aqueous solution, or the X-ray method. The “membrane” conformation is rare one and, to date has only been observed by NMR for a single cytotoxin 1 from N. oxiana (CT1No) in detergent micelle. Both CT13Nn and CT1No are S-type CTs. Membrane-binding of these CTs probably involves an additional step—the conformational transformation of the loop-2. To confirm this suggestion, we conducted molecular dynamics simulations of both CT1No and CT13Nn in the Highly Mimetic Membrane Model of palmitoiloleoylphosphat...
Biochemistry, 2008
Latarcins, linear peptides from the Lachesana tarabaevi spider venom, exhibit a broad-spectrum an... more Latarcins, linear peptides from the Lachesana tarabaevi spider venom, exhibit a broad-spectrum antimicrobial activity, likely acting on the bacterial cytoplasmic membrane. We study their spatial structures and interaction with model membranes by a combination of experimental and theoretical methods to reveal the structure-activity relationship. In this work, a 26 amino acid peptide, Ltc1, was investigated. Its spatial structure in detergent micelles was determined by (1)H nuclear magnetic resonance (NMR) and refined by Monte Carlo simulations in an implicit water-octanol slab. The Ltc1 molecule was found to form a straight uninterrupted amphiphilic helix comprising 8-23 residues. A dye-leakage fluorescent assay and (31)P NMR spectroscopy established that the peptide does not induce the release of fluorescent marker nor deteriorate the bilayer structure of the membranes. The voltage-clamp technique showed that Ltc1 induces the current fluctuations through planar membranes when the sign of the applied potential coincides with the one across the bacterial inner membrane. This implies that Ltc1 acts on the membranes via a specific mechanism, which is different from the carpet mode demonstrated by another latarcin, Ltc2a, featuring a helix-hinge-helix structure with a hydrophobicity gradient along the peptide chain. In contrast, the hydrophobic surface of the Ltc1 helix is narrow-shaped and extends with no gradient along the axis. We have also disclosed a number of peptides, structurally homologous to Ltc1 and exhibiting similar membrane activity. This indicates that the hydrophobic pattern of the Ltc1 helix and related antimicrobial peptides specifies their activity mechanism. The latter assumes the formation of variable-sized lesions, which depend upon the potential across the membrane.
Carditoxins (CTXs) from cobra snake venoms, the basic 60-62 residue allbeta sheet polypeptides, a... more Carditoxins (CTXs) from cobra snake venoms, the basic 60-62 residue allbeta sheet polypeptides, are known to bind to and impair the function of cell membranes. To assess the membrane induced conformation and orientation of CTXs, the interaction of the P-type cardiotoxin II from Naja oxiana snake venom (CTII) with perdeuterated dodecylphosphocholine (DPC) was studied using 1 H-NMR spectroscopy and diffusion measurements. Under conditions where the toxin formed a well-de®ned complex with DPC, the spatial structure of CTII with respect to the presence of tightly bound water molecules in loop II, was calculated using the torsion angle dynamics program DYANA. The structure was found to be similar, except for subtle changes in the tips of all three loops, to the previously described``major'' form of CTII in aqueous solution illustrated by thè`t rans'' con®guration of the Val7-Pro8 peptide bond. No``minor'' form with the``cis'' con®guration of the above bond was found in the micellebound state. The broadening of the CTII backbone proton signals by 5, 16-doxylstearate relaxation probes, together with modeling based on the spatial structure of CTII, indicated a periphery mode of binding of the toxin molecule to the micelle and revealed its micelle interacting domain. The latter includes a hydrophobic region of CTII within the extremities of loops I and III (residues 5-11, 46-50), the basement of loop II (residues 24-29,31-37) and the belt of polar residues encircling these loops (lysines 4,5,12,23,50, serines 11,46, histidine 31, arginine 36). It is suggested that this structural motif and the mode of binding can be realized during interaction of CTXs with lipid and biological membranes.
Biochemical and biophysical research communications, 2021
Cobra cytotoxins (CTs), the three-fingered proteins, feature high amino acid sequence homology in... more Cobra cytotoxins (CTs), the three-fingered proteins, feature high amino acid sequence homology in the beta-strands and variations in the loop regions. We selected a pair of cytotoxins from Naja kaouthia crude venom to clarify the sequence-structure relationships. Using chromatography and mass spectroscopy, we separated and identified the mixture of cytotoxins 2 and 3, differentiated by the only Val 41/Ala 41 substitution. Here, using natural abundance 13C, 15N NMR-spectroscopy we performed chemical shift assignments of the signals of the both toxins in aqueous solution in the major and minor forms. Combining NOE and chemical shift data, the toxins' spatial structure was determined. Finally, we proved that the tip of the "finger"-2, or the loop-2 of cytotoxins adopts the shape of an omega-loop with a tightly-bound water molecule in its cavity. Comparison with other NMR and X-ray structures of cytotoxins possessing different amino acid sequences reveals spatial similarit...
The pressure-induced changes in 15N enriched HPr fromStaphylococcus carnosus were investigated by... more The pressure-induced changes in 15N enriched HPr fromStaphylococcus carnosus were investigated by twodimensional~2D! heteronuclear NMR spectroscopy at pressures ranging from atmospheric pressure up to 200 MPa. The NMR experiments allowed the simultaneous observation of the backbone and side-chain amide protons and nitrogens. Most of the resonances shift downfield with increasing pressure indicating generalized pressure-induced conformational changes. The average pressure-induced shifts for amide protons and nitrogens are 0.285 ppm GPa 21 at 278 K and 2.20 ppm GPa 21, respectively. At 298 K the corresponding values are 0.275 and 2.41 ppm GPa 21. roton and nitrogen pressure coefficients show a significant but rather small correlation ~0.31! if determined for all amide resonances. When restricting the analysis to amide groups in the b-pleated sheet, the correlation between these coefficients is with 0.59 significantly higher. As already described for other proteins, the amide proton pr...
Journal of chemical information and modeling, 2021
For many peripheral membrane-binding polypeptides(MBPs), especially β-structural ones, the precis... more For many peripheral membrane-binding polypeptides(MBPs), especially β-structural ones, the precise molecular mechanisms of membrane insertion remain unclear. In most cases, only the terminal water-soluble and membrane-bound states have been elucidated, whereas potential functionally important intermediate stages are still not understood in sufficient detail. In this study, we present one of the first successful attempts to describe step-by-step embedding of the MBP cardiotoxin 2 (CT2) from cobra Naja oxiana venom into a lipid bilayer at the atomistic level. CT2 possesses a highly conservative and rigid β-structured three-finger fold shared by many other exogenous and endogenous proteins performing a wide variety of functions. The incorporation of CT2 into the lipid bilayer was analyzed via a 2 μs all-atom molecular dynamics (MD) simulation without restraints. This process was shown to occur over a number of distinct steps, while the geometry of initial membrane attachment drasticall...
Dynamics and folding of proteins are intimately related to their volumetric properties, such as v... more Dynamics and folding of proteins are intimately related to their volumetric properties, such as volume and compressibility associated with conformational states. These properties can only be explored in experiments in which pressure is a variable. Here, site-specific analysis of compressibility and stability of a protein is a major current concern from the viewpoint of structural biology. Combination of the on-line high pressure cell techniquewith a commercial high field NMR spectrometer (e.g., 750 MHz for proton) paves the way to the solution. Pressure-sensitive, flexible or fragile regions of a protein can be identified in this manner by 'H, 'tN and r3C NMR spectroscopy under varying hydrostatic pressure between I and-4 kbar. Pressure-induced changes of chemical shifts of essentially all the tH and r5N NMR signals along with changes in 'H nuclear Overhauser effect (NOE) reveal that the entire folded structure responds sensitively to pressure. However, the response can be quite nonuniform over the protein architecture, and there appear to be pressure-sensitive regions perhaps associated with low packing of atoms. The non-uniform structural response of a protein is a direct measure of non-uniform volume fluctuation within the folded ensemble. Pressure also affects directly dynamics of side chains in millisecond ranges, namely flip-flop motions of aromatic rings. Retardation of the flip motion at high pressure indicates that there is a fluctuation that produces a dynamic cavity in the protein core. In some proteins, denaturation occurs locally at certain segments of the protein at relatively low pressures (-1 kbar), followed by the unfolding of the entire polypeptide chain at higher pressures. It is likely that such a local denaturation may be initiated cavities of the protein, and may be considered to denaturation of proteins. by water penetration into hydrophobic be a general mechanism of Pressure
The following versions of software and data (see references i O) were used in the production of t... more The following versions of software and data (see references i O) were used in the production of this report:
Biochemical and Biophysical Research Communications
Toxins
In aqueous solutions, cobra cytotoxins (CTX), three-finger folded proteins, exhibit conformationa... more In aqueous solutions, cobra cytotoxins (CTX), three-finger folded proteins, exhibit conformational equilibrium between conformers with either cis or trans peptide bonds in the Nterminal loop (loop-I). The equilibrium is shifted to the cis form in toxins with a pair of adjacent Pro residues in this loop. It is known that CTX with a single Pro residue in loop-I and a cis peptide bond do not interact with lipid membranes. Thus, if a cis peptide bond is present in loop-I, as in a Pro-Pro containing CTX, this should weaken its lipid interactions and likely cytotoxic activities. To test this, we have isolated seven CTX from Naja naja and N. haje cobra venoms. Antibacterial and cytotoxic activities of these CTX, as well as their capability to induce calcein leakage from phospholipid liposomes, were evaluated. We have found that CTX with a Pro-Pro peptide bond indeed exhibit attenuated membrane-perturbing activity in model membranes and lower cytotoxic/antibacterial activity compared to their counterparts with a single Pro residue in loop-I.
Biochemistry (Moscow), 2014
Electrical charge and amphiphilicity are fundamental characteristics of biomolecules governing th... more Electrical charge and amphiphilicity are fundamental characteristics of biomolecules governing their activity in vivo. Amphiphilic peptides carrying a multitude of electrical charges are physiologically active due to their ability to interact with receptors and a variety of biomembrane components [1, 2]. In particular, polycationic peptides are considered as perspective molecules for antibacterial and anticancer therapy [3, 4]. Such peptides are widely present in the venoms of insects, arachnids, and snakes [5, 6]. Some of them, e.g. cardiotoxins, feature compact and highly stable spatial structure due to the presence of disulfide bonds [7, 8]. In contrast, linear peptides exhibit high flexibility and are typically able to switch among a number of states [9, 10]. In this work we studied amphiphilic peptides with high net positive charge-linear cationic peptides (LCP). These include latarcins Ltc1K [11] and Ltc2a [12] isolated from the venom of the Middle Asia Lachesana
Protein Expression and Purification, 2017
Cytotoxins or cardiotoxins is a group of polycationic toxins from cobra venom belonging to the &#... more Cytotoxins or cardiotoxins is a group of polycationic toxins from cobra venom belonging to the 'three-finger' protein superfamily (Ly6/uPAR family) which includes small β-structural proteins (60-90 residues) with high disulfide bond content (4-5 disulfides). Due to a high cytotoxic activity for cancer cells, cytotoxins are considered as potential anticancer agents. Development of the high-throughput production methods is required for the prospective applications of cytotoxins. Here, efficient approach for bacterial production of recombinant analogue of cytotoxin I from N. oxiana containing additional N-terminal Met-residue (rCTX1) was developed. rCTX1 was produced in the form of E. coli inclusion bodies. Refolding in optimized conditions provided ∼6 mg of correctly folded protein from 1 L of bacterial culture. Cytotoxicity of rCTX1 for C6 rat glioma cells was found to be similar to the activity of wild type CTX1. The milligram quantities of 13C,15N-labeled rCTX1 were obtained. NMR study confirmed the similarity of the spatial structures of recombinant and wild-type toxins. Additional Met residue does not perturb the overall structure of the three-finger core. The analysis of available data for different Ly6/uPAR proteins of snake and human origin revealed that efficiency of their folding in vitro is correlated with the number of proline residues in the third loop and the surface area of hydrophobic residues buried within the protein interior. The obtained data indicate that hydrophobic core is important for the folding of proteins with high disulfide bond content. Developed expression method opens new possibilities for structure-function studies of CTX1 and other related three-finger proteins.
Toxins, 2022
Cobra cytotoxins (CTs) belong to the three-fingered protein family and possess membrane activity.... more Cobra cytotoxins (CTs) belong to the three-fingered protein family and possess membrane activity. Here, we studied cytotoxin 13 from Naja naja cobra venom (CT13Nn). For the first time, a spatial model of CT13Nn with both “water” and “membrane” conformations of the central loop (loop-2) were determined by X-ray crystallography. The “water” conformation of the loop was frequently observed. It was similar to the structure of loop-2 of numerous CTs, determined by either NMR spectroscopy in aqueous solution, or the X-ray method. The “membrane” conformation is rare one and, to date has only been observed by NMR for a single cytotoxin 1 from N. oxiana (CT1No) in detergent micelle. Both CT13Nn and CT1No are S-type CTs. Membrane-binding of these CTs probably involves an additional step—the conformational transformation of the loop-2. To confirm this suggestion, we conducted molecular dynamics simulations of both CT1No and CT13Nn in the Highly Mimetic Membrane Model of palmitoiloleoylphosphat...
Biochemistry, 2008
Latarcins, linear peptides from the Lachesana tarabaevi spider venom, exhibit a broad-spectrum an... more Latarcins, linear peptides from the Lachesana tarabaevi spider venom, exhibit a broad-spectrum antimicrobial activity, likely acting on the bacterial cytoplasmic membrane. We study their spatial structures and interaction with model membranes by a combination of experimental and theoretical methods to reveal the structure-activity relationship. In this work, a 26 amino acid peptide, Ltc1, was investigated. Its spatial structure in detergent micelles was determined by (1)H nuclear magnetic resonance (NMR) and refined by Monte Carlo simulations in an implicit water-octanol slab. The Ltc1 molecule was found to form a straight uninterrupted amphiphilic helix comprising 8-23 residues. A dye-leakage fluorescent assay and (31)P NMR spectroscopy established that the peptide does not induce the release of fluorescent marker nor deteriorate the bilayer structure of the membranes. The voltage-clamp technique showed that Ltc1 induces the current fluctuations through planar membranes when the sign of the applied potential coincides with the one across the bacterial inner membrane. This implies that Ltc1 acts on the membranes via a specific mechanism, which is different from the carpet mode demonstrated by another latarcin, Ltc2a, featuring a helix-hinge-helix structure with a hydrophobicity gradient along the peptide chain. In contrast, the hydrophobic surface of the Ltc1 helix is narrow-shaped and extends with no gradient along the axis. We have also disclosed a number of peptides, structurally homologous to Ltc1 and exhibiting similar membrane activity. This indicates that the hydrophobic pattern of the Ltc1 helix and related antimicrobial peptides specifies their activity mechanism. The latter assumes the formation of variable-sized lesions, which depend upon the potential across the membrane.
Carditoxins (CTXs) from cobra snake venoms, the basic 60-62 residue allbeta sheet polypeptides, a... more Carditoxins (CTXs) from cobra snake venoms, the basic 60-62 residue allbeta sheet polypeptides, are known to bind to and impair the function of cell membranes. To assess the membrane induced conformation and orientation of CTXs, the interaction of the P-type cardiotoxin II from Naja oxiana snake venom (CTII) with perdeuterated dodecylphosphocholine (DPC) was studied using 1 H-NMR spectroscopy and diffusion measurements. Under conditions where the toxin formed a well-de®ned complex with DPC, the spatial structure of CTII with respect to the presence of tightly bound water molecules in loop II, was calculated using the torsion angle dynamics program DYANA. The structure was found to be similar, except for subtle changes in the tips of all three loops, to the previously described``major'' form of CTII in aqueous solution illustrated by thè`t rans'' con®guration of the Val7-Pro8 peptide bond. No``minor'' form with the``cis'' con®guration of the above bond was found in the micellebound state. The broadening of the CTII backbone proton signals by 5, 16-doxylstearate relaxation probes, together with modeling based on the spatial structure of CTII, indicated a periphery mode of binding of the toxin molecule to the micelle and revealed its micelle interacting domain. The latter includes a hydrophobic region of CTII within the extremities of loops I and III (residues 5-11, 46-50), the basement of loop II (residues 24-29,31-37) and the belt of polar residues encircling these loops (lysines 4,5,12,23,50, serines 11,46, histidine 31, arginine 36). It is suggested that this structural motif and the mode of binding can be realized during interaction of CTXs with lipid and biological membranes.
Biochemical and biophysical research communications, 2021
Cobra cytotoxins (CTs), the three-fingered proteins, feature high amino acid sequence homology in... more Cobra cytotoxins (CTs), the three-fingered proteins, feature high amino acid sequence homology in the beta-strands and variations in the loop regions. We selected a pair of cytotoxins from Naja kaouthia crude venom to clarify the sequence-structure relationships. Using chromatography and mass spectroscopy, we separated and identified the mixture of cytotoxins 2 and 3, differentiated by the only Val 41/Ala 41 substitution. Here, using natural abundance 13C, 15N NMR-spectroscopy we performed chemical shift assignments of the signals of the both toxins in aqueous solution in the major and minor forms. Combining NOE and chemical shift data, the toxins' spatial structure was determined. Finally, we proved that the tip of the "finger"-2, or the loop-2 of cytotoxins adopts the shape of an omega-loop with a tightly-bound water molecule in its cavity. Comparison with other NMR and X-ray structures of cytotoxins possessing different amino acid sequences reveals spatial similarit...
The pressure-induced changes in 15N enriched HPr fromStaphylococcus carnosus were investigated by... more The pressure-induced changes in 15N enriched HPr fromStaphylococcus carnosus were investigated by twodimensional~2D! heteronuclear NMR spectroscopy at pressures ranging from atmospheric pressure up to 200 MPa. The NMR experiments allowed the simultaneous observation of the backbone and side-chain amide protons and nitrogens. Most of the resonances shift downfield with increasing pressure indicating generalized pressure-induced conformational changes. The average pressure-induced shifts for amide protons and nitrogens are 0.285 ppm GPa 21 at 278 K and 2.20 ppm GPa 21, respectively. At 298 K the corresponding values are 0.275 and 2.41 ppm GPa 21. roton and nitrogen pressure coefficients show a significant but rather small correlation ~0.31! if determined for all amide resonances. When restricting the analysis to amide groups in the b-pleated sheet, the correlation between these coefficients is with 0.59 significantly higher. As already described for other proteins, the amide proton pr...
Journal of chemical information and modeling, 2021
For many peripheral membrane-binding polypeptides(MBPs), especially β-structural ones, the precis... more For many peripheral membrane-binding polypeptides(MBPs), especially β-structural ones, the precise molecular mechanisms of membrane insertion remain unclear. In most cases, only the terminal water-soluble and membrane-bound states have been elucidated, whereas potential functionally important intermediate stages are still not understood in sufficient detail. In this study, we present one of the first successful attempts to describe step-by-step embedding of the MBP cardiotoxin 2 (CT2) from cobra Naja oxiana venom into a lipid bilayer at the atomistic level. CT2 possesses a highly conservative and rigid β-structured three-finger fold shared by many other exogenous and endogenous proteins performing a wide variety of functions. The incorporation of CT2 into the lipid bilayer was analyzed via a 2 μs all-atom molecular dynamics (MD) simulation without restraints. This process was shown to occur over a number of distinct steps, while the geometry of initial membrane attachment drasticall...
Dynamics and folding of proteins are intimately related to their volumetric properties, such as v... more Dynamics and folding of proteins are intimately related to their volumetric properties, such as volume and compressibility associated with conformational states. These properties can only be explored in experiments in which pressure is a variable. Here, site-specific analysis of compressibility and stability of a protein is a major current concern from the viewpoint of structural biology. Combination of the on-line high pressure cell techniquewith a commercial high field NMR spectrometer (e.g., 750 MHz for proton) paves the way to the solution. Pressure-sensitive, flexible or fragile regions of a protein can be identified in this manner by 'H, 'tN and r3C NMR spectroscopy under varying hydrostatic pressure between I and-4 kbar. Pressure-induced changes of chemical shifts of essentially all the tH and r5N NMR signals along with changes in 'H nuclear Overhauser effect (NOE) reveal that the entire folded structure responds sensitively to pressure. However, the response can be quite nonuniform over the protein architecture, and there appear to be pressure-sensitive regions perhaps associated with low packing of atoms. The non-uniform structural response of a protein is a direct measure of non-uniform volume fluctuation within the folded ensemble. Pressure also affects directly dynamics of side chains in millisecond ranges, namely flip-flop motions of aromatic rings. Retardation of the flip motion at high pressure indicates that there is a fluctuation that produces a dynamic cavity in the protein core. In some proteins, denaturation occurs locally at certain segments of the protein at relatively low pressures (-1 kbar), followed by the unfolding of the entire polypeptide chain at higher pressures. It is likely that such a local denaturation may be initiated cavities of the protein, and may be considered to denaturation of proteins. by water penetration into hydrophobic be a general mechanism of Pressure
The following versions of software and data (see references i O) were used in the production of t... more The following versions of software and data (see references i O) were used in the production of this report:
Biochemical and Biophysical Research Communications