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Ekaterina Kolesanova

Donetsk National University, Laboratory of Peptide Engineering, Head of Laboratory

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Born November, 14, 1956.Graduated from school June, 1974 (Donetsk, USSR)Graduated from the Donetsk State University, Chemical Department, Biochemistry in June, 1979.PhD courses at MM Shemyakin Institute of Bioorganic Chemistry, USSR Academy of Sciences, Moscow, USSR -Dec 1979 - Dec 1982.Junior Scientist at the Institute of Physiologically Active Substances, Chernogolovka, Moscow region, USSR Dec 1982-Jan 1986.Research scietist at the All-Union Centre for Medico-Biological Problems of Narcology, Moscow, USSR Feb 1986 - June 1991Senior research scientist, leadins scientist, Head of Laboratory at the Institute of Biomedical Chemistry, Moscow, Russia 1991-now
Address: Russian Federation

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Papers by Ekaterina Kolesanova

Influence of amino acid and N-terminal protection residue structures on peptide p-nitroanilide adsorption on polystyrene-based support

Amino Acids, Jul 14, 2023

Influence of amino acid and N-terminal protection residue structures on peptide p-nitroanilide adsorption on polystyrene-based support

Research Square (Research Square), May 19, 2023

[Antigenic mapping of cytochrome P450 101 (P450cam)]

Bioorganicheskaia khimiia, 1998

Eighteen linear antigenic sites were found in cytochrome P450 101 (P450cam) from Pseudomonas puti... more Eighteen linear antigenic sites were found in cytochrome P450 101 (P450cam) from Pseudomonas putida by the peptide scanning method. These sites accounted for about 30% of the protein sequence. We found no sequences that completely coincided with the antigenic sites of P450cam in cytochromes P450 from other sources. The linear B-epitopes of P540cam were mainly localized on the boundaries separating the elements of the secondary structure. Seventeen of eighteen antigenic sites were found to be on the protein surface and accessible to water molecules. Many functionally important sites or amino acid residues of the P450cam molecules coincided or were in close proximity to the linear B-epitopes found.

Epitope mapping of horseradish peroxidase (isoenzyme C)

Biochemistry. Biokhimii͡a, 1997

Peptide scanning (PEPSCAN) was used to determine linear antigenic determinants of horseradish per... more Peptide scanning (PEPSCAN) was used to determine linear antigenic determinants of horseradish peroxidase isoenzyme C (HRPC). For this purpose, we synthesized 303 overlapping hexapeptide fragments (with a step of one amino acid residue) of the protein primary structure and studied their interactions with anti-HRPC polyclonal antisera by ELISA. Experiments with various titers of antisera allowed us to determine linear antigenic determinants of HRPC; several such determinants were spatially located in regular elements of the secondary structure (alpha-helices) found both inside and outside the protein globule. A fraction of epitopes were located in loops and folds of the HRPC peptide chain with irregular shapes. These epitopes contained several functionally important residues: Arg 38, which is part of the active site of the enzyme, as well as Phe 142 and Phe 143, which form a channel allowing aromatic substrates to reach the active site. Amino acid residues that form calcium-binding si...

B-Cell Linear Epitopes in the Conservative Regions of Hepatitis C Virus Envelope Glycoproteins

Peptides: The Wave of the Future, 2001

Hepatitis C virus (HCV) infection in 80% cases causes chronic hepatitis C development, often resu... more Hepatitis C virus (HCV) infection in 80% cases causes chronic hepatitis C development, often resulting in cirrhosis and hepatocarcinonaa. High variability of HCV El and E2 envelope protein fragments, to which virus-neutralizing antibodies are formed in the natural course of infection, enables the virus to escape from the host immune response. Nevertheless, El and E2 sequences contain highly conservative sites that may be responsible for all attachment and entry and may elicit protective antibodies [1–3]. However, these sites have not been defined and their immunogenicity has not been subjected to regular studies.

Development of the schedule for multiple parallel "difficult" Peptide synthesis on pins

International journal of peptides, 2013

Unified schedule for multiple parallel solid-phase synthesis of so-called "difficult" p... more Unified schedule for multiple parallel solid-phase synthesis of so-called "difficult" peptides on polypropylene pins was developed. Increase in the efficiency of 9-fluorenyl(methoxycarbonyl) N-terminal amino-protecting group removal was shown to have a greater influence on the accuracy of the "difficult" peptide synthesis than the use of more efficient amino acid coupling reagents such as aminium salts. Hence the unified schedule for multiple parallel solid-phase synthesis of "difficult" peptides included the procedure for N-terminal amino group deprotection modified by applying a more efficient reagent for the deprotection and the standard procedure of amino acid coupling by carbodiimide method with an additional coupling using aminium salts, if necessary. Amino acid coupling with the help of carbodiimide allows to follow the completeness of the coupling via the bromophenol blue indication, thus providing the accuracy of the synthesis and preventing an...

Bacteriophage MS2 As a Tool for Targeted Delivery in Solid Tumor Chemotherapy

Acta Naturae

Bacteriophage MS2 was employed for targeted delivery of an apoptosis-inducing agent, Tl+, into a ... more Bacteriophage MS2 was employed for targeted delivery of an apoptosis-inducing agent, Tl+, into a tumor tissue. The targeted delivery was ensured by iRGD peptide, a ligand of integrins presumably located on the surface of endotheliocytes of the tumor tissue neovasculature and certain tumor cells. The synthesized peptide was conjugated to MS2 capsid proteins. Tl+ ions from TlNO3 penetrated the phage particles and tightly bound to phage RNA. Peptide-modified MS2 preparations filled with Tl+ caused cell death in two types of cultivated human breast cancer cells and effected necrosis of these tumor xenografts in mice. Neither peptide-conjugated bacteriophage MS2 without Tl+ nor the phage filled with Tl+ but without the peptide or the same phage with the non-conjugated peptide in solution produced such effects. The preparation exhibited no acute toxicity at a therapeutic dose.

Urine nerve growth factor in patients with Idiopathic detrusor overctivity and in patients with overactive bladder without detrusor overactivity

Study design, materials and methods 21 patient with idiopathic DO (IDO) and18 patients with OAB w... more Study design, materials and methods 21 patient with idiopathic DO (IDO) and18 patients with OAB without DO were enrolled. All of them had OAB symptoms: urgency, frequency and urgent urinary incontinence. During the examination all patients were performed the urodynamic study.11volonteers without lower urinary tract symptoms were also enrolled in the control group. For measurement of urinary NGF level we used the ELISA method. In our studies, we use undiluted voiding urine that is placed on ice immediately and centrifuged at 3000 g at 4°C for 10 minutes. The supernatant is separated into aliquots in 1.5-mL tubes and preserved in a 80°C freezer. At the same time, 3 mL of urine is taken to measure urinary creatinine level. We determine urinary NGF concentration using specific ELISA kit that has a minimum sensitivity of 7.8 pg/mL. The amount of NGF that falls below the detection limits of NGF assay are extracted from an NGF standard curve. The total urinary NGF level is further normaliz...

Peptides specifically target bacteriophage MS2, filled with an apoptosis-inducingagent, to tumour

Proceedings of the 35th European Peptide Symposium

Synthesis of peptides glycated at Lys residues

Proceedings of the 35th European Peptide Symposium

Epitope mapping of cytochrome P450 2B4 by peptide scanning

Biochemistry and molecular biology international

Overlapping hexapeptides covering the whole sequence of the cytochrome P450 2B4 have been synthes... more Overlapping hexapeptides covering the whole sequence of the cytochrome P450 2B4 have been synthesized on the solid supports and tested by ELISA using the polyclonal antiserum against cytochrome 2B4. 70 hexapeptide fragments have been found to interact specifically with the antiserum, i.e. to possess antigenic activity. The mapped linear epitopes occupy about 43% of the whole sequence of 2B4. They presumably form clusters in the regions of No. 60-150, 210-300, 390-430 and 465-486 amino acid residues. The use of cytochrome P450 DataBase has allowed to classify the revealed antigenic determinants into absolutely specific for 2B4, specific only for 2B4 and 2B5, characteristic for 2B subfamily and widely distributed in family 2.

Comparison of Reagents for Fmoc-Group Removal from Growing Peptide Chain in the Solid-Phase Peptide Synthesis

Journal of Peptide Science

To avoid problems arising from the use of the precursor piperidine as a reagent for Fmoc-group re... more To avoid problems arising from the use of the precursor piperidine as a reagent for Fmoc-group removal in solid-phase peptide synthesis (SPPS) several secondary amines were tested as substitutes of piperidine in this reaction. Kinetics of Fmoc removal as well as manual SPPS yields and by-product formation were compared with piperidine, piperazine, morpholine, pyrrolidine, 4-methyl-piperidine and also their mixtures with DBU used as Fmoc removing reagents. While possessing the same kinetic parameters of Fmoc removal as piperidine, 4-methyl-piperidine was shown to be superior to all tested reagents in terms of the desired product yields. The use of 4-methyl-piperidine in automated SPPS demonstrated its equal efficiency to piperidine in the synthesis process and superiority to piperidine in its behavior in liquid handling modules, i.e., a complete absence of tubing pellets. It makes 4- methyl-piperidine a good substitute for piperidine with no restrictions relevant to the latter as a p...

Spontaneous Charge Generation in Flowing Albumin Solutions at 35 °C and 38 °C

Biosensors, Jan 11, 2017

The time dependence of a charge accumulation in a 10-15 M albumin solution, flowing through a mea... more The time dependence of a charge accumulation in a 10-15 M albumin solution, flowing through a measuring cell of an analytical flow system injector, had a nonlinear character under certain conditions, within a human physiological temperature range. Sharp charge increases depended on albumin concentration. This effect must be taken into consideration when generating models that describe electrokinetic phenomena in flowing protein solutions and when developing analytical flow systems for the registration of biomolecules in low concentration ranges.

Computer Design of Vaccines: Approaches, Software Tools and Informational Resources

by Ekaterina Kolesanova and Vladimir Poroikov

Current Computer - Aided Drug Design, 2005

Enkephalin metabolism in rat brain after bestatin administration

Peptide Science — Present and Future, 2002

[Dynamics of met-enkephalin level in the adrenal glands and plasma of rats during acute alcoholic intoxication]

Voprosy medit͡sinskoĭ khimii

Content of Met-enkephalin in rat adrenal glands and blood plasma as well as activity of the enzym... more Content of Met-enkephalin in rat adrenal glands and blood plasma as well as activity of the enzymatic systems catalyzing the enkephalins catabolism in blood serum were studied after a single administration of ethanol into rats. Dynamics of Met-enkephalin alterations studied suggest that phases of activation and inhibition occur in the secretion of the peptide from adrenal gland into circulation.

[Comparative characteristics of soluble and membrane brain aminopeptidases. I. Isolation, physico-chemical properties, catalytic activity]

Bioorganicheskaia khimiia

The methods were worked out for isolating of the bovine brain soluble and membrane-bound aminopep... more The methods were worked out for isolating of the bovine brain soluble and membrane-bound aminopeptidases in highly purified state. One of the stages involved a biospecific-type chromatography on aminohexyl-Sepharose. Both enzymes were found to have equal molecular masses (ca. 100-107 kD) and isoelectric points (pI 4,6). None of the enzymes possessed a subunit structure. Both aminopeptidases were inactivated by omicron-phenanthroline and by an SH-reagent, p-hydroxymercuribenzoate. The catalytic constants for the hydrolysis of a specific substrate, L-leucine p-nitroanilide, were identical for the two enzymes. So far no differences in the physico-chemical or enzymatic properties of the soluble and membrane-bound enzymes were disclosed.

[Comparative characteristics of soluble and membrane brain aminopeptidases. II. Substrate specificity]

Bioorganicheskaia khimiia

The methods were worked out for isolating of the bovine brain soluble and membrane-bound aminopep... more The methods were worked out for isolating of the bovine brain soluble and membrane-bound aminopeptidases in highly purified state. One of the stages involved a biospecific-type chromatography on aminohexyl-Sepharose. Both enzymes were found to have equal molecular masses (ca. 100-107 kD) and isoelectric points (pI 4,6). None of the enzymes possessed a subunit structure. Both aminopeptidases were inactivated by omicron-phenanthroline and by an SH-reagent, p-hydroxymercuribenzoate. The catalytic constants for the hydrolysis of a specific substrate, L-leucine p-nitroanilide, were identical for the two enzymes. So far no differences in the physico-chemical or enzymatic properties of the soluble and membrane-bound enzymes were disclosed.

[The role of single amino acid residues of the immunodominant continuous epitope of cytochrome P450CAM (CYP101) 312LKKGDQ317 in recognition by specific antibodies]

Voprosy medit͡sinskoĭ khimii

As it has been shown previously the site 311-318 of bacterial cytochrome P450cam (CYP101) contain... more As it has been shown previously the site 311-318 of bacterial cytochrome P450cam (CYP101) contains an immunodominant continuous B-epitope. In order to investigate the role of single amino acid residues in antibody binding antigenic hexapeptide 312LKKGDQ317 analogues including single amino acid replacements were synthesized using Multipin technology. Antibodies from three anti-P450cam polyclonal rabbit sera interacted similarly to these peptides. The residue G315 was found to play a significant role in antibody recognition; any replacement leads there to considerable decrease of antibody binding. Residues L312, K313 and D316 occurred to be partly replaceable, whereas K314 and Q317 were not essential in recognition. These results correspond to known spatial structure of P450cam molecule. In its 312-317 site the polypeptide chain makes a turn and so some water-accessible atoms form a compact surface cluster including side chain atoms of K313 and D316, O-atom of K314 and C alpha-atom of G315, which present reliable Ig binding site. Side chain of K314 outlying from this cluster does not participate in the interaction. The received data permit to consider that the continuous epitope 312-317 of P450cam is conformationally dependent.

[Aminopeptidases and their inhibitors]

Vestnik Akademii meditsinskikh nauk SSSR

Influence of amino acid and N-terminal protection residue structures on peptide p-nitroanilide adsorption on polystyrene-based support

Amino Acids, Jul 14, 2023

Influence of amino acid and N-terminal protection residue structures on peptide p-nitroanilide adsorption on polystyrene-based support

Research Square (Research Square), May 19, 2023

[Antigenic mapping of cytochrome P450 101 (P450cam)]

Bioorganicheskaia khimiia, 1998

Eighteen linear antigenic sites were found in cytochrome P450 101 (P450cam) from Pseudomonas puti... more Eighteen linear antigenic sites were found in cytochrome P450 101 (P450cam) from Pseudomonas putida by the peptide scanning method. These sites accounted for about 30% of the protein sequence. We found no sequences that completely coincided with the antigenic sites of P450cam in cytochromes P450 from other sources. The linear B-epitopes of P540cam were mainly localized on the boundaries separating the elements of the secondary structure. Seventeen of eighteen antigenic sites were found to be on the protein surface and accessible to water molecules. Many functionally important sites or amino acid residues of the P450cam molecules coincided or were in close proximity to the linear B-epitopes found.

Epitope mapping of horseradish peroxidase (isoenzyme C)

Biochemistry. Biokhimii͡a, 1997

Peptide scanning (PEPSCAN) was used to determine linear antigenic determinants of horseradish per... more Peptide scanning (PEPSCAN) was used to determine linear antigenic determinants of horseradish peroxidase isoenzyme C (HRPC). For this purpose, we synthesized 303 overlapping hexapeptide fragments (with a step of one amino acid residue) of the protein primary structure and studied their interactions with anti-HRPC polyclonal antisera by ELISA. Experiments with various titers of antisera allowed us to determine linear antigenic determinants of HRPC; several such determinants were spatially located in regular elements of the secondary structure (alpha-helices) found both inside and outside the protein globule. A fraction of epitopes were located in loops and folds of the HRPC peptide chain with irregular shapes. These epitopes contained several functionally important residues: Arg 38, which is part of the active site of the enzyme, as well as Phe 142 and Phe 143, which form a channel allowing aromatic substrates to reach the active site. Amino acid residues that form calcium-binding si...

B-Cell Linear Epitopes in the Conservative Regions of Hepatitis C Virus Envelope Glycoproteins

Peptides: The Wave of the Future, 2001

Hepatitis C virus (HCV) infection in 80% cases causes chronic hepatitis C development, often resu... more Hepatitis C virus (HCV) infection in 80% cases causes chronic hepatitis C development, often resulting in cirrhosis and hepatocarcinonaa. High variability of HCV El and E2 envelope protein fragments, to which virus-neutralizing antibodies are formed in the natural course of infection, enables the virus to escape from the host immune response. Nevertheless, El and E2 sequences contain highly conservative sites that may be responsible for all attachment and entry and may elicit protective antibodies [1–3]. However, these sites have not been defined and their immunogenicity has not been subjected to regular studies.

Development of the schedule for multiple parallel "difficult" Peptide synthesis on pins

International journal of peptides, 2013

Unified schedule for multiple parallel solid-phase synthesis of so-called "difficult" p... more Unified schedule for multiple parallel solid-phase synthesis of so-called "difficult" peptides on polypropylene pins was developed. Increase in the efficiency of 9-fluorenyl(methoxycarbonyl) N-terminal amino-protecting group removal was shown to have a greater influence on the accuracy of the "difficult" peptide synthesis than the use of more efficient amino acid coupling reagents such as aminium salts. Hence the unified schedule for multiple parallel solid-phase synthesis of "difficult" peptides included the procedure for N-terminal amino group deprotection modified by applying a more efficient reagent for the deprotection and the standard procedure of amino acid coupling by carbodiimide method with an additional coupling using aminium salts, if necessary. Amino acid coupling with the help of carbodiimide allows to follow the completeness of the coupling via the bromophenol blue indication, thus providing the accuracy of the synthesis and preventing an...

Bacteriophage MS2 As a Tool for Targeted Delivery in Solid Tumor Chemotherapy

Acta Naturae

Bacteriophage MS2 was employed for targeted delivery of an apoptosis-inducing agent, Tl+, into a ... more Bacteriophage MS2 was employed for targeted delivery of an apoptosis-inducing agent, Tl+, into a tumor tissue. The targeted delivery was ensured by iRGD peptide, a ligand of integrins presumably located on the surface of endotheliocytes of the tumor tissue neovasculature and certain tumor cells. The synthesized peptide was conjugated to MS2 capsid proteins. Tl+ ions from TlNO3 penetrated the phage particles and tightly bound to phage RNA. Peptide-modified MS2 preparations filled with Tl+ caused cell death in two types of cultivated human breast cancer cells and effected necrosis of these tumor xenografts in mice. Neither peptide-conjugated bacteriophage MS2 without Tl+ nor the phage filled with Tl+ but without the peptide or the same phage with the non-conjugated peptide in solution produced such effects. The preparation exhibited no acute toxicity at a therapeutic dose.

Urine nerve growth factor in patients with Idiopathic detrusor overctivity and in patients with overactive bladder without detrusor overactivity

Study design, materials and methods 21 patient with idiopathic DO (IDO) and18 patients with OAB w... more Study design, materials and methods 21 patient with idiopathic DO (IDO) and18 patients with OAB without DO were enrolled. All of them had OAB symptoms: urgency, frequency and urgent urinary incontinence. During the examination all patients were performed the urodynamic study.11volonteers without lower urinary tract symptoms were also enrolled in the control group. For measurement of urinary NGF level we used the ELISA method. In our studies, we use undiluted voiding urine that is placed on ice immediately and centrifuged at 3000 g at 4°C for 10 minutes. The supernatant is separated into aliquots in 1.5-mL tubes and preserved in a 80°C freezer. At the same time, 3 mL of urine is taken to measure urinary creatinine level. We determine urinary NGF concentration using specific ELISA kit that has a minimum sensitivity of 7.8 pg/mL. The amount of NGF that falls below the detection limits of NGF assay are extracted from an NGF standard curve. The total urinary NGF level is further normaliz...

Peptides specifically target bacteriophage MS2, filled with an apoptosis-inducingagent, to tumour

Proceedings of the 35th European Peptide Symposium

Synthesis of peptides glycated at Lys residues

Proceedings of the 35th European Peptide Symposium

Epitope mapping of cytochrome P450 2B4 by peptide scanning

Biochemistry and molecular biology international

Overlapping hexapeptides covering the whole sequence of the cytochrome P450 2B4 have been synthes... more Overlapping hexapeptides covering the whole sequence of the cytochrome P450 2B4 have been synthesized on the solid supports and tested by ELISA using the polyclonal antiserum against cytochrome 2B4. 70 hexapeptide fragments have been found to interact specifically with the antiserum, i.e. to possess antigenic activity. The mapped linear epitopes occupy about 43% of the whole sequence of 2B4. They presumably form clusters in the regions of No. 60-150, 210-300, 390-430 and 465-486 amino acid residues. The use of cytochrome P450 DataBase has allowed to classify the revealed antigenic determinants into absolutely specific for 2B4, specific only for 2B4 and 2B5, characteristic for 2B subfamily and widely distributed in family 2.

Comparison of Reagents for Fmoc-Group Removal from Growing Peptide Chain in the Solid-Phase Peptide Synthesis

Journal of Peptide Science

To avoid problems arising from the use of the precursor piperidine as a reagent for Fmoc-group re... more To avoid problems arising from the use of the precursor piperidine as a reagent for Fmoc-group removal in solid-phase peptide synthesis (SPPS) several secondary amines were tested as substitutes of piperidine in this reaction. Kinetics of Fmoc removal as well as manual SPPS yields and by-product formation were compared with piperidine, piperazine, morpholine, pyrrolidine, 4-methyl-piperidine and also their mixtures with DBU used as Fmoc removing reagents. While possessing the same kinetic parameters of Fmoc removal as piperidine, 4-methyl-piperidine was shown to be superior to all tested reagents in terms of the desired product yields. The use of 4-methyl-piperidine in automated SPPS demonstrated its equal efficiency to piperidine in the synthesis process and superiority to piperidine in its behavior in liquid handling modules, i.e., a complete absence of tubing pellets. It makes 4- methyl-piperidine a good substitute for piperidine with no restrictions relevant to the latter as a p...

Spontaneous Charge Generation in Flowing Albumin Solutions at 35 °C and 38 °C

Biosensors, Jan 11, 2017

The time dependence of a charge accumulation in a 10-15 M albumin solution, flowing through a mea... more The time dependence of a charge accumulation in a 10-15 M albumin solution, flowing through a measuring cell of an analytical flow system injector, had a nonlinear character under certain conditions, within a human physiological temperature range. Sharp charge increases depended on albumin concentration. This effect must be taken into consideration when generating models that describe electrokinetic phenomena in flowing protein solutions and when developing analytical flow systems for the registration of biomolecules in low concentration ranges.

Computer Design of Vaccines: Approaches, Software Tools and Informational Resources

by Ekaterina Kolesanova and Vladimir Poroikov

Current Computer - Aided Drug Design, 2005

Enkephalin metabolism in rat brain after bestatin administration

Peptide Science — Present and Future, 2002

[Dynamics of met-enkephalin level in the adrenal glands and plasma of rats during acute alcoholic intoxication]

Voprosy medit͡sinskoĭ khimii

Content of Met-enkephalin in rat adrenal glands and blood plasma as well as activity of the enzym... more Content of Met-enkephalin in rat adrenal glands and blood plasma as well as activity of the enzymatic systems catalyzing the enkephalins catabolism in blood serum were studied after a single administration of ethanol into rats. Dynamics of Met-enkephalin alterations studied suggest that phases of activation and inhibition occur in the secretion of the peptide from adrenal gland into circulation.

[Comparative characteristics of soluble and membrane brain aminopeptidases. I. Isolation, physico-chemical properties, catalytic activity]

Bioorganicheskaia khimiia

The methods were worked out for isolating of the bovine brain soluble and membrane-bound aminopep... more The methods were worked out for isolating of the bovine brain soluble and membrane-bound aminopeptidases in highly purified state. One of the stages involved a biospecific-type chromatography on aminohexyl-Sepharose. Both enzymes were found to have equal molecular masses (ca. 100-107 kD) and isoelectric points (pI 4,6). None of the enzymes possessed a subunit structure. Both aminopeptidases were inactivated by omicron-phenanthroline and by an SH-reagent, p-hydroxymercuribenzoate. The catalytic constants for the hydrolysis of a specific substrate, L-leucine p-nitroanilide, were identical for the two enzymes. So far no differences in the physico-chemical or enzymatic properties of the soluble and membrane-bound enzymes were disclosed.

[Comparative characteristics of soluble and membrane brain aminopeptidases. II. Substrate specificity]

Bioorganicheskaia khimiia

The methods were worked out for isolating of the bovine brain soluble and membrane-bound aminopep... more The methods were worked out for isolating of the bovine brain soluble and membrane-bound aminopeptidases in highly purified state. One of the stages involved a biospecific-type chromatography on aminohexyl-Sepharose. Both enzymes were found to have equal molecular masses (ca. 100-107 kD) and isoelectric points (pI 4,6). None of the enzymes possessed a subunit structure. Both aminopeptidases were inactivated by omicron-phenanthroline and by an SH-reagent, p-hydroxymercuribenzoate. The catalytic constants for the hydrolysis of a specific substrate, L-leucine p-nitroanilide, were identical for the two enzymes. So far no differences in the physico-chemical or enzymatic properties of the soluble and membrane-bound enzymes were disclosed.

[The role of single amino acid residues of the immunodominant continuous epitope of cytochrome P450CAM (CYP101) 312LKKGDQ317 in recognition by specific antibodies]

Voprosy medit͡sinskoĭ khimii

As it has been shown previously the site 311-318 of bacterial cytochrome P450cam (CYP101) contain... more As it has been shown previously the site 311-318 of bacterial cytochrome P450cam (CYP101) contains an immunodominant continuous B-epitope. In order to investigate the role of single amino acid residues in antibody binding antigenic hexapeptide 312LKKGDQ317 analogues including single amino acid replacements were synthesized using Multipin technology. Antibodies from three anti-P450cam polyclonal rabbit sera interacted similarly to these peptides. The residue G315 was found to play a significant role in antibody recognition; any replacement leads there to considerable decrease of antibody binding. Residues L312, K313 and D316 occurred to be partly replaceable, whereas K314 and Q317 were not essential in recognition. These results correspond to known spatial structure of P450cam molecule. In its 312-317 site the polypeptide chain makes a turn and so some water-accessible atoms form a compact surface cluster including side chain atoms of K313 and D316, O-atom of K314 and C alpha-atom of G315, which present reliable Ig binding site. Side chain of K314 outlying from this cluster does not participate in the interaction. The received data permit to consider that the continuous epitope 312-317 of P450cam is conformationally dependent.

[Aminopeptidases and their inhibitors]

Vestnik Akademii meditsinskikh nauk SSSR