TY - JOUR

T1 - Altai maral (Cervus elaphus sibiricus) chymosin structural characterization and modeling the enzyme interaction with the chymosin-sensitive regions of κ-caseins

AU - Belenkaya, S V

AU - M G, Ilyina

AU - Borisevich, S S

AU - Khamitov, E M

AU - Diusenova, S E

AU - Shevtsov, M B

AU - Borshchevskiy, V I

AU - Kolosov, P V

AU - Fomin, A S

AU - Arkhipov, S G

AU - Volosnikova, E A

AU - Elchaninov, V V

AU - Kolybalov, D S

AU - Shcherbakov, D N

N1 - This work was supported by the Ministry of Science and Higher Education of the Russian Federation within the governmental order for SRF “SKIF” Boreskov Institute of Catalysis (FWUR-2024-0040). Сrystallographic data collection was supported by the Ministry of Science and Higher Education of the Russian Federation (agreement 075-03-2025-662, project FSMG-2025-0012)., Altai maral (Cervus elaphus sibiricus) chymosin structural characterization and modeling the enzyme interaction with the chymosin-sensitive regions of κ-caseins / S. V. Belenkaya, M. G. Ilyina, S. S. Borisevich, E. M. Khamitov, S. E. Diusenova, M.B. Shevtsov, V. I. Borshchevskiy, P. V. Kolosov, A. S. Fomin, S. G. Arkhipov, E. A. Volosnikova, V. V. Elchaninov, D. S. Kolybalov, D. N. Shcherbakov // International Journal of Biological Macromolecules. - 2025. - № 330. Ч. 4. - С. 148085.

PY - 2025/10/3

Y1 - 2025/10/3

N2 - Chymosins (Chn's) are not only popular industrial milk coagulants, but also attractive models for studying the mechanisms of target peptide bonds recognition by aspartate peptidases. Despite the high amino acid sequence homology and similarity of their tertiary structures, Chns of various mammals may have significantly different enzymatic properties. We have obtained Altai maral (Cervus elaphus sibiricus) crystal structure. Bovine and dromedary camel chymosins with known structures and characterized specificities were used for comparative analysis. The recombinant maral Chn (rChn-Cer) specificity was determined by chromatograph mass spectrometry after enzyme incubation with peptide substrates repeating the maral, bovine and camel κ-caseines (κ-CN's) chymosine sensitive regions. Analysis of resulting products indicates that in the maral and bovine κ-CN sequences, maral rChn hydrolyzes the F105-M106 bond, but in the camel substrate, it hydrolyzes the F97-I98 bond. According to the comparative analysis of spatial structures, the arrangement of catalytic amino acid residues of maral, bovine and camel Chn's is almost identical. The obtained rChn-cer structure were used as a starting model for protein-protein docking and molecular dynamics to describe the interactions of the enzyme catalytic center with synthetic peptide substrates repeating the chymosine sensitive regions of maral, bovine and camel κ-CN. The obtained results could be useful for understanding the mechanisms of interaction of chymosins with substrates of different species.

AB - Chymosins (Chn's) are not only popular industrial milk coagulants, but also attractive models for studying the mechanisms of target peptide bonds recognition by aspartate peptidases. Despite the high amino acid sequence homology and similarity of their tertiary structures, Chns of various mammals may have significantly different enzymatic properties. We have obtained Altai maral (Cervus elaphus sibiricus) crystal structure. Bovine and dromedary camel chymosins with known structures and characterized specificities were used for comparative analysis. The recombinant maral Chn (rChn-Cer) specificity was determined by chromatograph mass spectrometry after enzyme incubation with peptide substrates repeating the maral, bovine and camel κ-caseines (κ-CN's) chymosine sensitive regions. Analysis of resulting products indicates that in the maral and bovine κ-CN sequences, maral rChn hydrolyzes the F105-M106 bond, but in the camel substrate, it hydrolyzes the F97-I98 bond. According to the comparative analysis of spatial structures, the arrangement of catalytic amino acid residues of maral, bovine and camel Chn's is almost identical. The obtained rChn-cer structure were used as a starting model for protein-protein docking and molecular dynamics to describe the interactions of the enzyme catalytic center with synthetic peptide substrates repeating the chymosine sensitive regions of maral, bovine and camel κ-CN. The obtained results could be useful for understanding the mechanisms of interaction of chymosins with substrates of different species.

UR - https://pubmed.ncbi.nlm.nih.gov/41046897/

UR - https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=105018860964&origin=inward

U2 - 10.1016/j.ijbiomac.2025.148085

DO - 10.1016/j.ijbiomac.2025.148085

M3 - Article

C2 - 41046897

VL - 330

SP - 148085

JO - International Journal of Biological Macromolecules

JF - International Journal of Biological Macromolecules

SN - 1879-0003

IS - Pt 4

ER -