猪链球菌3-羟基-3-甲基戊二酰辅酶A还原酶的原核表达、活性检测和同源建模

doi:10.11843/j.issn.0366-6964.2016.11.021

Abstract

Abstract:

The aim of this study was to take the Streptococcus suis (S. suis) 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) as a drug target, and provide theoretical and experimental basis for the development of novel antibiotics for the prevention and treatment of S. suis infection. In this experiment, S. suis HMGR was prokaryotically expressed and purified by Ni-NTA agrose. Its enzyme activity was determined by spectrophotometric method. In addition, a multiple sequence alignment of HMGR was performed by CLUSTALW to identify conserved domains. Homology modeling of the three-dimensional (3D) structure of S. suis HMGR was performed by SWISS-MODEL. The quality of the model was evaluated by PROCHECK. The result of enzyme activity analyses showed that optimal reaction conditions of S. suis HMGR was pH 5.0 and 37 ℃, the Vmax and Km value was 846 U•mg^-1 and 0.213 mmol•L^－1 respectively. The analysis of SWISS-MODEL showed that crystal structure of Streptococcus pneumonia HMGR was the appropriate template for homology modeling of S. suis HMGR. The theoretic model of 3D structure of S. suis HMGR was proven to be reliable through quality assessment. The S. suis HMGR model will contribute to virtual screening of novel antibacterial agents against S. suis infection. Enzyme activity assay provides a feasible method for validation of effective inhibitors against S. suis HMGR in biochemical experiment.

CLC Number:

S852.61

LIU Can-ying, XU Zhuo-fei, FU Qiang. Prokaryotic Expression, Activity Assay and Homology Modeling of the 3-hydroxy-3-methylglutaryl Coenzyme A Reductase from Streptococcus suis[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(11): 2318-2324.

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References

［1］WINDSOR R S, ELLIOTT S D. Streptococcal infection in young pigs. IV. An outbreak of streptococcal meningitis in weaned pigs［J］. J Hyg (Lond), 1975, 75(1): 69-78.
［2］WINDSOR R S. Meningitis in pigs caused by Streptococcus suis type 2［J］. Vet Rec, 1977, 101（19）: 378-379.
［3］ERICKSON E D. Streptococcosis［J］. J Am Vet Med Assoc, 1987, 191（11）: 1391-1393.
［4］HIGGINS R, GOTTSCHALK M, MITTAL K R, et al. Streptococcus suis infection in swine. A sixteen month study［J］. Can J Vet Res, 1990, 54（1）: 170-173.
［5］CHANTER N, JONES P W, ALEXANDER T J. Meningitis in pigs caused by Streptococcus suis-a speculative review［J］. Vet Microbiol, 1993, 36（1-2）: 39-55.
［6］GOYETTE-DESJARDINS G, AUGER J P, XU J, et al. Streptococcus suis, an important pig pathogen and emerging zoonotic agent-an update on the worldwide distribution based on serotyping and sequence typing［J］. Emerg Microbes Infect, 2014, 3 (6): e45.
［7］WERTHEIM H F, NGUYEN H N, TAYLOR W, et al. Streptococcus suis, an important cause of adult bacterial meningitis in northern Vietnam［J］. PLoS One, 2009, 4(6):e5973.
［8］REAMS R Y, GLICKMAN L T, HARRINGTON D D, et al. Streptococcus suis infection in swine: a retrospective study of 256 cases. Part I. Epidemiologic factors and antibiotic susceptibility patterns［J］. J Vet Diag Invest, 1993, 5（3）: 363-367.
［9］HEUSTON S, BEGLEY M, GAHAN C G, et al. Isoprenoid biosynthesis in bacterial pathogens［J］. Microbiology, 2012, 158(Pt6):1389-1401.
［10］REUSCH V M Jr. Lipopolymers, isoprenoids, and the assembly of the gram-positive cell wall［J］. Crit Rev Microbiol, 1984, 11（2）:129-155.
［11］MATSUMI R, ATOMI H, DRIESSEN A J, et al. Isoprenoid biosynthesis in Archaea-biochemical and evolutionary implications［J］. Res Microbiol, 2011, 162（1）: 39-52.
［12］BOUCHER Y, DOOLITTLE W F. The role of lateral gene transfer in the evolution of isoprenoid biosynthesis pathways［J］. Mol Microbiol, 2000, 37(4): 703-716.
［13］SETO H, WATANABE H, FURIHATA K. Simultaneous operation of the mevalonate and non-mevalonate pathways in the biosynthesis of isopentenyl diphosphate in Streptomyces aeriouvifer［J］. Tetrahedron Lett, 1996, 37（44）: 7979-7982.
［14］WILDING E I, BROWN J R, BRYANT A P, et al. Identification, evolution, and essentiality of the mevalonate pathway for isopentenyl diphosphate biosynthesis in gram-positive cocci［J］. J Bacteriol, 2000, 182（15）:4319-4327.
［15］WILDING E I, KIM D Y, BRYANT A P, et al. Essentiality, expression, and characterization of the Class II 3-hydroxy-3-methylglutaryl coenzymeA reductase of Staphylococcus aureus［J］. J Bacteriol, 2000, 182(18): 5147-5152.
［16］BOCHAR D A, STAUFFACHER C V, RODWELL V W. Sequence comparisons reveal two classes of 3-hydroxy-3-methylglutaryl coenzymeA reductase［J］. Mol Genet Metab, 1999, 66(2):122-127.
［17］MIZIORKO H M. Enzymes of the mevalonate pathway of isoprenoid biosynthesis［J］. Arch Biochem Biophys, 2011, 505(2): 131-143.
［18］LAWRENCE C M, RODWELL V W, STAUFFACHER C V. Crystalstructure of Pseudomonas mevalonii HMG-CoA reductase at 3.0 angstrom resolution［J］. Science, 1995, 268(5218):1758-1762.
［19］ISTVAN E S. Bacterial and mammalian HMG-CoA reductases: related enzymes with distinct architectures［J］. Curr Opin Struct Biol, 2001, 11(6): 746-751.
［20］ALBERTS A W, CHEN J, KURON G, et al. Mevinolin: a highly potent competitive inhibitor of hydroxymethylglutaryl-coenzyme A reductase and a cholesterol-lowering agent［J］. Proc Natl Acad Sci U S A, 1980, 77(7): 3957-3961.
［21］LI W, HU X Y, LIU L, et al. Induction of protective immune response against Streptococcus suis serotype 2 infection by the surface antigen HP0245［J］. FEMS Microbiol Lett, 2011, 316(2): 115-122.
［22］李鹏, 陈兰英. 血脂康抑制猪肝HMG-CoA还原酶的活力［J］. 基础医学与临床, 2003, 23(5): 531-534.
LI P, CHEN L Y. Xuezhikang inhibits the activity of HMG-CoA reductase in pig liver［J］. Basic Medical Sciences and Clinics, 2003, 23(5): 531-534.（in Chinese）
［23］LI K B. ClustalW-MPI: ClustalW analysis using distributed and parallel computing［J］. Bioinformatics, 2003, 19(12): 1585-1586.
［24］BIASINI M, BIENERT S, WATERHOUSE A, et al. SWISS-MODEL: modelling protein tertiary and quaternary structure using evolutionary information［J］. Nucleic Acids Res, 2014, 42: 252-258.
［25］LASKOWSKI R A, MACARTHUR M W, MOSS D S, et al. PROCHECK: a program to check the stereochemical quality of protein structures［J］. J Appl Cryst, 1993, 26(2): 283-291.
［26］FENG Y J, ZHANG H M, WU Z W, et al. Streptococcus suis infection an emerging/reemerging challenge of bacterial infectious diseases?［J］. Virulence, 2014, 5(4): 477-497
［27］LI D, GUI J, LI Y J, et al. Structure-based design and screen of novel inhibitors for class II 3-hydroxy-3-methylglutaryl coenzyme A reductase from Streptococcus pneumoniae［J］. J Chem Inf Model, 2012, 52(7): 1833-1841.
［28］BEN Y L, CUI G Z, LI C, et al. Expression, purification, characteristics and homology modeling of the HMGS from Streptococcus penumoniae［J］. Biomed Environ Sci, 2009, 22(3): 229-236.

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Prokaryotic Expression, Activity Assay and Homology Modeling of the 3-hydroxy-3-methylglutaryl Coenzyme A Reductase from Streptococcus suis

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