pH-regulated Secretion of a Glyceraldehyde-3-Phosphate Dehydrogenase from Streptococcus gordonii FSS2: Purification,Characterization,and Cloning of the Gene Encoding this Enzyme
Restricted accessOtherFirst published online January, 2001
pH-regulated Secretion of a Glyceraldehyde-3-Phosphate Dehydrogenase from Streptococcus gordonii FSS2: Purification,Characterization,and Cloning of the Gene Encoding this Enzyme
Streptococcus gordonii and other viridans streptococci (VS) are primary etiologic agents of infective endocarditis, despite being part of the normal oral microflora. Recently, a surface-bound glyceraldehyde-3 -phosphate dehydrogenase (GAPDH) has been found on the cells of all tested streptococcal species, where it has been implicated as a virulence factor. In contrast, we observed that a soluble extracellular GAPDH was the major secreted protein from S. gordonii FSS2, an endocarditis strain. The biochemical properties and gene sequence of S. gordonii GAPDH are almost identical to those of other streptococcal GAPDHs. Growth at defined pHs showed that secretion of GAPDH is regulated by environmental pH. GAPDH was primarily surface-associated at growth pH 6.5 and shifted to > 90% secreted at growth pH 7.5. Others have identified S. gordonii promoters that are up-regulated by a pH shift similar to that experienced by organisms entering the blood stream (neutral) from the oral cavity (slightly acid). Analysis of our results suggests that secretion of GAPDH may be a similar adaptation by S. gordonii.
Bibby BG, Mundorff SA, Zero DT, Almekinder KJ (1986). Oral food clearance and the pH of plaque and saliva . J Am Dent Assoc112:333-337.
2.
Boyd DA, Cvitkovitch DG, Hamilton IR (1995). Sequence, expression, and function of the gene for the nonphosphorylating, NADPdependent glyceraldehyde-3-phosphate dehydrogenase of Streptococcus mutans. J Bacteriol177:2622-2627.
3.
Bradshaw DJ, Marsh PD (1998). Analysis of pH-driven disruption of oral microbial communities in vitro. Caries Res32:456-462.
4.
Brown AT, Wittenberger CL (1971). The occurrence of multiple glyceraldehyde-3-phosphate dehydrogenases in cariogenic streptococci. Biochem Biophys Res Commun43:217-224.
5.
Cancilla MR, Hillier AJ, Davidson BE (1995). Lactococcus lactis glyceraldehyde-3-phosphate dehydrogenase gene, gap: further evidence for strongly biased codon usage in glycolytic pathway genes. Microbiology141:1027-1036.
6.
Dashper SG, Reynolds EC (2000). Effects of organic acid anions on growth, glycolysis, and intracellular pH of oral streptococci. J Dent Res79:90-96.
7.
Durack DT (1990). Infective and noninfective endocarditis. In: The heart, arteries and veins. Hurst JW, Schlant RC, Rackley CF, Sonnenblick EH , Wenger NK, editors. New York: McGraw-Hill, pp. 1230-1255.
8.
Eichenbaum Z., Green BD, Scott JR (1996). Iron starvation causes release from the group A streptococcus of the ADP-ribosylating protein called plasmin receptor or surface glyceraldehyde-3-phosphate dehydrogenase. Infect Immun64:1956-1960.
9.
Ferdinand W. (1964). The isolation and specific activity of rabbit-muscle glyceraldehyde phosphate dehydrogenase. Biochem J92:578-585.
10.
Gase K., Gase A., Schirmer H., Malke H. (1996). Cloning, sequencing and functional overexpression of the Streptococcus equisimilis H46A gapC gene encoding a glyceraldehyde-3-phosphate dehydrogenase that also functions as a plasmin(ogen)-binding protein. Purification and biochemical characterization of the protein. Eur J Biochem239:42-51.
11.
Harris JI, Hocking JD, Runswick MJ, Suzuki K., Walker JE (1980). D-glyceraldehyde-3-phosphate dehydrogenase. The purification and characterization of the enzyme from the thermophiles Bacillus stearothermophilus and Thermus aquaticus. Eur J Biochem108:535-547.
12.
Harty Dws, Mayo JA, Cook SL, Jacques NA (2000). Environmental regulation of glycosidase and peptidase production by Streptococcus gordonii FSS2. Microbiology146:1923-1931.
13.
Herzberg MC (1996). Platelet-streptococcal interactions in endocarditis . Crit Rev Oral Biol Med7:222-236.
14.
Herzberg MC, Meyer MW, Kiliç A., Tao L. (1997). Host-pathogen interactions in bacterial endocarditis: streptococcal virulence in the host. Adv Dent Res11:69-74.
15.
Horecker BL, Kornberg A. (1948). The extinction coefficient of the reduced band of pyridine nucleotides. J Biol Chem175:385-390. Kiliç AO, Herzberg MC, Meyer MW, Zhao X., Tao L. (1999). Streptococcal reporter gene-fusion vector for identification of in vivo expressed genes. Plasmid42:67-72.
16.
Kolberg J., Sletten K. (1996). Monoclonal antibodies that recognize a common pneumococcal protein with similarities to streptococcal group A surface glyceraldehyde-3-phosphate dehydrogenase. Infect Immun64:3544-3547.
17.
Korzeniowski OM, Kaye D. ( 1992). Infective endocarditis. In: Heart disease; a textbook of cardiovascular medicine. Braunwald E, editor. Philadelphia: W.B. Saunders Company, pp. 1078-1105.
18.
Krebs EG (1955). Glyceraldehyde-3-phosphate dehydrogenase from yeast. In: Methods in enzymology. Colowick SP, Kaplan NO, editors. New York: Academic Press, Inc. , pp. 407-411.
19.
Lamkin GE, King EE ( 1976). Blue sepharose: a reusable affinity chromatography medium for purification of alcohol dehydrogenase. Biochem Biophys Res Commun72:560-565.
20.
Lee SF (1992). Identification and characterization of a surface protein-releasing activity in Streptococcus mutans and other pathogenic streptococci . Infect Immun60:4032-4039.
21.
Lee SF (1995). Active release of bound antibody by Streptococcus mutans. Infect Immun63:1940-1946.
22.
Loesche WJ (1986). Role of Streptococcus mutans in human dental decay . Microbiol Rev50:353-380.
23.
Lottenberg R., Broder CC, Boyle MD, Kain SJ, Schroeder BL, Curtiss R Iii (1992a). Cloning, sequence analysis, and expression in Escherichia coli of a streptococcal plasmin receptor. J Bacteriol174:5204-5210.
24.
Lottenberg R., DesJardin LE, Wang H., Boyle MD (1992b). Streptokinase-producing streptococci grown in human plasma acquire unregulated cell-associated plasmin activity. J Infect Dis166:436-440.
25.
Manning JE, Hume EB, Hunter N., Knox KW ( 1994). An appraisal of the virulence factors associated with streptococcal endocarditis. J Med Microbiol40: 110-114.
26.
Matsudaira P. (1987). Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes. J Biol Chem262:10035-10038.
27.
Mayo JA, Zhu H., Harty Dws, Knox KW ( 1995). Modulation of glycosidase and protease activities by chemostat growth conditions in an endocarditis strain of Streptococcus sanguis . Oral Microbiol Immunol10:342-348.
28.
Navarre WW, Schneewind O (1999). Surface proteins of Gram-positive bacteria and mechanisms of their targeting to the cell wall envelope. Microbiol Mol Biol Rev63:174-229.
29.
Pancholi V., Fischetti VA (1992). A major surface protein on group A streptococci is a glyceraldehyde-3-phosphate dehydrogenase with multiple binding activity . J Exp Med176:415-426.
30.
Pancholi V., Fischetti VA (1993). Glyceraldehyde-3-phosphate dehydrogenase on the surface of group A streptococci is also an ADP-ribosylating enzyme. Proc Natl Acad Sci USA90:8154-8158.
31.
Pancholi V., Fischetti VA (1997a). Cell-to-cell signaling between group A streptococci and pharyngeal cells. In: Streptococci and the host. Horaud T, Bouvet A, Leclercq R, de Montcos H , Sicard M, editors. New York: Plenum Press, pp. 499-504.
32.
Pancholi V., Fischetti VA (1997b). Regulation of the phosphorylation of human pharyngeal cell proteins by group A streptococcal surface dehydrogenase: signal transduction between streptococci and pharyngeal cells. J Exp Med186:1633-1643.
33.
Schagger H., von Jagow G. (1987). Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem166:368-379.
34.
Sirover MA (1999). New insights into an old protein: the functional diversity of mammalian glyceraldehyde-3-phosphate dehydrogenase. Biochim Biophys Acta1432:159-184.
35.
Sneath Pha, Mair NS, Sharpe ME (1984). Bergey's manual of systematic bacteriology. Baltimore: Williams and Wilkins, pp. 1043-1071.
36.
van Houte J. (1994). Role of micro-organisms in caries etiology. J Dent Res73:672-681.
37.
Velick SF (1955). Glyceraldehyde-3-phosphate dehydrogenase from muscle. In: Methods in enzymology. Colowick SP, Kaplan NO, editors. New York: Academic Press, Inc. , pp. 401-406.
38.
Vriesema Ajm, Dankert J., Zaat Saj (2000a). A shift from oral to blood pH is a stimulus for adaptive gene expression of Streptococcus gordonii CHI and induces protection against oxidative stress and enhanced bacterial growth by expression of msrA . Infect Immun68:1061-1068.
39.
Vriesema Ajm, Brinkman R., Kok J., Dankert J., Zaat Saj (2000b). Broad-host-range shuttle vectors for screening of regulated promoter activity in viridans group streptococci: isolation of a pH-regulated promoter. Appl Environ Microbiol66:535-542.
40.
Whiley RA, Beighton D. (1998). Current classification of the oral streptococci . Oral Microbiol Immunol13:195-216.
41.
Winram SB, Lottenberg R. (1996). The plasmin-binding protein Plr of group A streptococci is identified as glyceraldehyde-3-phosphate dehydrogenase. Microbiology142:2311-2320.
