Link between sortase A function and cariogenicity of Streptococcus mutans: a preliminary metabolomics analysis

doi:10.7518/hxkq.2018.04.003

Abstract

Abstract:

Objective This study intends to explore the mechanism underlying the support of sortase A (SrtA) of the cario-genicity of Streptococcus mutans (S. mutans). Methods We performed a metabonomics study based on ¹H nuclear magnetic resonance spectroscopy (NMR), in which we compared the extracellular metabolites of wild-type S. mutans UA159 with those of its SrtA-deficient strain. Metabolite differences among strains were identified using a combination of principal component analysis and orthogonality partial least square discriminant analysis. Results Several differences corresponding mostly to unknown metabolites were identified. Some amino acids such as leucine and valine (δ 0.92×10^-6-1.20×10^-6), lactic acid ( δ1.28×10^-6), oxoglutaric acid (δ 3.00×10^-6), and glycine (δ 3.60×10^-6) differed among strains. Conclusion This work establishes the feasibility of using ¹H NMR-based metabonomics to provide leads for research into molecular factors that promote caries. The database of microbial metabolites should be also improved in further studies.

Key words: metabonomics, Streptococcus mutans, sortase A, ¹H nuclear magnetic resonance spectroscopy, princi-pal component analysis, orthogonality partial least squares discrimination analysis

CLC Number:

R780.2

Ying Song, Jinglin Zhou, Yuanli He, Wei Li, Ling Zou. Link between sortase A function and cariogenicity of Streptococcus mutans: a preliminary metabolomics analysis[J]. West China Journal of Stomatology, 2018, 36(4): 360-366.

Figures/Tables 7

Fig 1

Fig 2

Fig 3

Fig 4

Fig 5

Fig 6

Tab 1

References 48

[1]	Kuramitsu HK, Wang BY.The whole is greater than the sum of its parts: dental plaque bacterial interactions can affect the virulence properties of cariogenic Streptococcus mutans[J]. Am J Dent, 2011, 24(3): 153-154.
[2]	Bowen WH, Koo H.Biology of Streptococcus mutans-derived glucosyltransferases: role in extracellular matrix formation of cariogenic biofilms[J]. Caries Res, 2011, 45(1): 69-86.
[3]	Aoki H, Shiroza T, Hayakawa M, et al.Cloning of a Strep-tococcus mutans glucosyltransferase gene coding for inso-luble glucan synthesis[J]. Infect Immun, 1986, 53(3): 587-594.
[4]	Pucci MJ, Jones KR, Kuramitsu HK, et al.Molecular cloning and characterization of the glucosyltransferase C gene (gtfC) from Streptococcus mutans LM7[J]. Infect Immun, 1987, 55(9): 2176-2182.
[5]	Hanada N, Kuramitsu HK.Isolation and characterization of the Streptococcus mutans gtfD gene, coding for primer-dependent soluble glucan synthesis[J]. Infect Immun, 1989, 57(7): 2079-2085.
[6]	Inagaki S, Fujita K, Takashima Y, et al.Regulation of recom-bination between gtfB/gtfC genes in Streptococcus mutans by recombinase A[J]. Sci World J, 2013, 2013: 405075.
[7]	Okahashi N, Sasakawa C, Yoshikawa M, et al.Molecular characterization of a surface protein antigen gene from sero-type c Streptococcus mutans, implicated in dental caries[J]. Mol Microbiol, 1989, 3(5): 673-678.
[8]	Igarashi T, Asaga E, Goto N.The sortase of Streptococcus mutans mediates cell wall anchoring of a surface protein antigen[J]. Oral Microbiol Immunol, 2003, 18(4): 266-269.
[9]	Brady LJ, Maddocks SE, Larson MR, et al.The changing faces of Streptococcus antigen Ⅰ/Ⅱ polypeptide family adhesins[J]. Mol Microbiol, 2010, 77(2): 276-286.
[10]	Banas JA, Potvin HC, Singh RN.The regulation of Strep-tococcus mutans glucan-binding protein A expression[J]. FEMS Microbiol Lett, 1997, 154(2): 289-292.
[11]	Sato Y, Yamamoto Y, Kizaki H.Cloning and sequence ana-lysis of the gbpC gene encoding a novel glucan-binding protein of Streptococcus mutans[J]. Infect Immun, 1997, 65(2): 668-675.
[12]	Mattos-Graner RO, Jin S, King WF, et al.Cloning of the Streptococcus mutans gene encoding glucan binding protein B and analysis of genetic diversity and protein production in clinical isolates[J]. Infect Immun, 2001, 69(11): 6931-6941.
[13]	Shah DS, Russell RR.A novel glucan-binding protein with lipase activity from the oral pathogen Streptococcus mutans[J]. Microbiology, 2004, 150(Pt 6): 1947-1956.
[14]	Han T, Zhang C, Dao M.Identification and characteriza-tion of collagen-binding activity in Streptococcus mutans wall-associated protein: a possible implication in dental root caries and endocarditis[J]. Biochem Biophys Res Commun, 2006, 343(3): 787-792.
[15]	Igarashi T, Asaga E, Sato Y, et al.Inactivation of srtA gene of Streptococcus mutans inhibits dextran-dependent aggre-gation by glucan-binding protein C[J]. Oral Microbiol Im-munol, 2004, 19(1): 57-60.
[16]	Zhu L, Kreth J, Cross SE, et al.Functional characterization of cell-wall-associated protein WapA in Streptococcus mu-tans[J]. Microbiology, 2006, 152(Pt 8): 2395-2404.
[17]	Wen ZT, Burne RA.Analysis of cis- and trans-acting factors involved in regulation of the Streptococcus mutans fructa-nase gene (fruA)[J]. J Bacteriol, 2002, 184(1): 126-133.
[18]	Igarashi T, Asaga E, Goto N.Roles of Streptococcus mutans dextranase anchored to the cell wall by sortase[J]. Oral Mi-crobiol Immunol, 2004, 19(2): 102-105.
[19]	Igarashi T.Deletion in sortase gene of Streptococcus mutans Ingbritt[J]. Oral Microbiol Immunol, 2004, 19(3): 210-213.
[20]	Lee SF, McGavin MK. Identification of a point mutation resulting in loss of cell wall anchoring activity of SrtA of Streptococcus mutans NG5[J]. Infect Immun, 2004, 72(7): 4314-4317.
[21]	Lee SF, Boran TL.Roles of sortase in surface expression of the major protein adhesin P1, saliva-induced aggregation and adherence, and cariogenicity of Streptococcus mutans[J]. Infect Immun, 2003, 71(2): 676-681.
[22]	Lévesque CM, Voronejskaia E, Huang YC, et al.Involve-ment of sortase anchoring of cell wall proteins in biofilm formation by Streptococcus mutans[J]. Infect Immun, 2005, 73(6): 3773-3777.
[23]	姜颖, 张铁柱, 杨锦波, 等. 葡萄糖浓度对变形链球菌srtA基因表达影响的研究[J]. 口腔医学, 2009, 29(9): 461-464.
	Jiang Y, Zhang TZ, Yang JB, et al.Effects of glucose con-centrations on Streptococcus mutans srtA expression[J]. Sto-matology, 2009, 29(9): 461-464.
[24]	姜颖, 张铁柱, 杨锦波, 等. 酸性环境对变形链球菌srtA基因表达的影响[J]. 现代口腔医学杂志, 2009, 23(3): 286-288.
	Jiang Y, Zhang TZ, Yang JB, et al.Effect of acidic environ-ment on Streptococcus mutans srtA expression[J]. J Modern Stomatol, 2009, 23(3): 286-288.
[25]	Yang ZG, Nakabayashi R, Okazaki Y, et al.Toward better annotation in plant metabolomics: isolation and structure elucidation of 36 specialized metabolites from Oryza sativa (rice) by using MS/MS and NMR analyses[J]. Metabolomics, 2014, 10(4): 543-555.
[26]	Jordan KW, Nordenstam J, Lauwers GY, et al.Metabolomic characterization of human rectal adenocarcinoma with intact tissue magnetic resonance spectroscopy[J]. Dis Colon Rec-tum, 2009, 52(3): 520-525.
[27]	Bu Q, Huang YN, Yan GY, et al.Metabolomics: a revolu-tion for novel cancer marker identification[J]. Comb Chem High Throughput Screen, 2012, 15(3): 266-275.
[28]	Daviss B.Growing pains for metabolomics[J]. Scientist, 2005, 19(8): 25-28.
[29]	Zhang JT, Zhang Y, Du YY, et al.Dynamic metabonomic responses of tobacco (Nicotiana tabacum) plants to salt stress[J]. J Proteome Res, 2011, 10(4): 1904-1914.
[30]	Liu CX, Hao FH, Hu J, et al.Revealing different systems responses to brown planthopper infestation for pest suscep-tible and resistant rice plants with the combined metabonomic and gene-expression analysis[J]. J Proteome Res, 2010, 9(12): 6774-6785.
[31]	Chiang KP, Niessen S, Saghatelian A, et al.An enzyme that regulates ether lipid signaling pathways in cancer annotated by multidimensional profiling[J]. Chem Biol, 2006, 13(10): 1041-1050.
[32]	Zhao X, Huang CY, Lei HH, et al.Dynamic metabolic res-ponse of mice to acute mequindox exposure[J]. J Proteome Res, 2011, 10(11): 5183-5190.
[33]	Robertson DG.Metabonomics in toxicology: a review[J]. Toxicol Sci, 2005, 85(2): 809-822.
[34]	Buchholz A, Hurlebaus J, Wandrey C, et al.Metabolomics: quantification of intracellular metabolite dynamics[J]. Bio-mol Eng, 2002, 19(1): 5-15.
[35]	Wu JF, Xu WX, Ming ZP, et al.Metabolic changes reveal the development of schistosomiasis in mice[J]. PLoS Negl Trop Dis, 2010, 4(8): e807.
[36]	Gibney MJ, Walsh M, Brennan L, et al.Metabolomics in human nutrition: opportunities and challenges[J]. Am J Clin Nutr, 2005, 82(3): 497-503.
[37]	Zhou JL, Xu B, Huang J, et al.1H NMR-based metabonomic and pattern recognition analysis for detection of oral squa-mous cell carcinoma[J]. Clin Chim Acta, 2009, 401(1/2): 8-13.
[38]	Nascimento MM, Gordan VV, Garvan CW, et al.Correla-tions of oral bacterial arginine and urea catabolism with caries experience[J]. Oral Microbiol Immunol, 2009, 24(2): 89-95.
[39]	Aas JA, Griffen AL, Dardis SR, et al.Bacteria of dental caries in primary and permanent teeth in children and young adults[J]. J Clin Microbiol, 2008, 46(4): 1407-1417.
[40]	Liao SM, Klein MI, Heim KP, et al.Streptococcus mutans extracellular DNA is upregulated during growth in biofilms, actively released via membrane vesicles, and influenced by components of the protein secretion machinery[J]. J Bac-teriol, 2014, 196(13): 2355-2366.
[41]	Schauer N, Steinhauser D, Strelkov S, et al.GC-MS libraries for the rapid identification of metabolites in complex biolo-gical samples[J]. FEBS Lett, 2005, 579(6): 1332-1337.
[42]	Gika HG, Theodoridis GA, Wingate JE, et al.Within-day reproducibility of an HPLC-MS-based method for metabo-nomic analysis: application to human urine[J]. J Proteome Res, 2007, 6(8): 3291-3303.
[43]	Soga T, Ohashi Y, Ueno Y, et al.Quantitative metabolome analysis using capillary electrophoresis mass spectrometry[J]. J Proteome Res, 2003, 2(5): 488-494.
[44]	Beckonert O, Keun HC, Ebbels TM, et al.Metabolic pro-filing, metabolomic and metabonomic procedures for NMR spectroscopy of urine, plasma, serum and tissue extracts[J]. Nat Protoc, 2007, 2(11): 2692-2703.
[45]	Nicholson JK, Lindon JC.Systems biology: metabonomics[J]. Nature, 2008, 455(7216): 1054-1056.
[46]	Griffin JL.Metabonomics: NMR spectroscopy and pattern recognition analysis of body fluids and tissues for charac-terisation of xenobiotic toxicity and disease diagnosis[J]. Curr Opin Chem Biol, 2003, 7(5): 648-654.
[47]	Grivet JP, Delort AM, Portais JC.NMR and microbiology: from physiology to metabolomics[J]. Biochimie, 2003, 85(9): 823-840.
[48]	Williams RE, Lenz EM, Evans JA, et al.A combined 1H NMR and HPLC-MS-based metabonomic study of urine from obese (fa/fa) Zucker and normal Wistar-derived rats[J]. J Pharm Biomed Anal, 2005, 38(3): 465-471.

代谢产物	变量/×10^-6	VIP
亮氨酸和缬氨酸	0.96	1.534 33
	0.92	1.319 41
	1.08	1.306 00
乳酸	1.28	1.236 70
甘氨酸	3.60	1.078 84
酮戊二酸	3.00	1.347 58