title: Functional mechanism of C-terminal tail in the enzymatic role of porcine testicular carbonyl reductase: a combined experiment and molecular dynamics simulation study of the C-terminal tail in the enzymatic role of PTCR.
abstract: Porcine testicular carbonyl reductase, PTCR which is one of the short chain dehydrogenases/reductases (SDR) superfamily catalyzes the NADPH-dependent reduction of carbonyl compounds including steroids and prostaglandins. Previously we reported C-terminal tail of PTCR was deleted due to a nonsynonymous single nucleotide variation (nsSNV). Here we identified from kinetic studies that the enzymatic properties for 5α-dihydrotestosterone (5α-DHT) were different between wild-type and C-terminal-deleted PTCRs. Compared to wild-type PTCR, C-terminal-deleted PTCR has much higher reduction rate. To investigate structural difference between wild-type and C-terminal-deleted PTCRs upon 5α-DHT binding, we performed molecular dynamics simulations for two complexes. Using trajectories, molecular interactions including hydrogen bonding patterns, distance between 5α-DHT and catalytic Tyr193, and interaction energies are analyzed and compared. During the MD simulation time, the dynamic behavior of C-terminal tail in wild-type PTCR is also examined using essential dynamics analysis. The results of our simulations reveal that the binding conformation of 5α-DHT in C-terminal-deleted PTCR is more favorable for reduction reaction in PTCR, which shows strong agreement with kinetic data. These structural findings provide valuable information to understand substrate specificity of PTCR and further kinetic properties of enzymes belonging to the SDR superfamily.
authors.affiliation: Division of Applied Life Science (BK21 Plus Program), Systems and Synthetic Agrobiotech Center (SSAC), Plant Molecular Biology and Biotechnology Research Center (PMBBRC), Research Institute of Natural Science (RINS), Gyeongsang National University (GNU), 501 Jinju-daero, Gazha-dong, Jinju, Republic of Korea.
authors.forename: Minky
authors.initials: M
authors.lastname: Son
authors.affiliation: Industry-Academic Cooperation Foundation, Gyeongnam National University of Science & Technology, Jinju, Republic of Korea.
authors.forename: Woo Young
authors.initials: WY
authors.lastname: Bang
authors.affiliation: Division of Applied Life Science (BK21 Plus Program), Systems and Synthetic Agrobiotech Center (SSAC), Plant Molecular Biology and Biotechnology Research Center (PMBBRC), Research Institute of Natural Science (RINS), Gyeongsang National University (GNU), 501 Jinju-daero, Gazha-dong, Jinju, Republic of Korea.
authors.forename: Chanin
authors.initials: C
authors.lastname: Park
authors.affiliation: Division of Applied Life Science (BK21 Plus Program), Systems and Synthetic Agrobiotech Center (SSAC), Plant Molecular Biology and Biotechnology Research Center (PMBBRC), Research Institute of Natural Science (RINS), Gyeongsang National University (GNU), 501 Jinju-daero, Gazha-dong, Jinju, Republic of Korea.
authors.forename: Yuno
authors.initials: Y
authors.lastname: Lee
authors.affiliation: Swine Science and Technology Center, Gyeongnam National University of Science & Technology, Jinju, Republic of Korea.
authors.forename: Seul Gi
authors.initials: SG
authors.lastname: Kwon
authors.affiliation: Swine Science and Technology Center, Gyeongnam National University of Science & Technology, Jinju, Republic of Korea.
authors.forename: Sam Woong
authors.initials: SW
authors.lastname: Kim
authors.affiliation: Swine Science and Technology Center, Gyeongnam National University of Science & Technology, Jinju, Republic of Korea.
authors.forename: Chul Wook
authors.initials: CW
authors.lastname: Kim
authors.affiliation: Division of Applied Life Science (BK21 Plus Program), Systems and Synthetic Agrobiotech Center (SSAC), Plant Molecular Biology and Biotechnology Research Center (PMBBRC), Research Institute of Natural Science (RINS), Gyeongsang National University (GNU), 501 Jinju-daero, Gazha-dong, Jinju, Republic of Korea.
authors.forename: Keun Woo
authors.initials: KW
authors.lastname: Lee
mesh_terms: D000449:Aldehyde Reductase / Q000737:chemistry* / Q000235:genetics; D000074408:Aldo-Keto Reductases; D000818:Animals; D055162:Biocatalysis; D020134:Catalytic Domain; D013196:Dihydrotestosterone / Q000737:chemistry*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D007700:Kinetics; D008297:Male; D062105:Molecular Docking Simulation; D056004:Molecular Dynamics Simulation; D009249:NADP / Q000737:chemistry*; D010084:Oxidation-Reduction; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000235:genetics; D013379:Substrate Specificity; D013552:Swine; D013737:Testis / Q000737:chemistry / Q000201:enzymology; D013816:Thermodynamics
chemical_list: D011993:Recombinant Fusion Proteins; D013196:Dihydrotestosterone; D009249:NADP; D000074408:Aldo-Keto Reductases; D000449:Aldehyde Reductase