The Biotin-dependent Sodium Ion Pump Glutaconyl-CoA Decarboxylase from Fusobacterium Nucleatum (subsp. Nucleatum). Comparison with the Glutaconyl-CoA Decarboxylases from Gram-positive Bacteria

Overview

Journal Arch Microbiol

Specialty Microbiology

Date 1990 Jan 1

PMID 2244788

Citations 16

Authors

B Beatrix

K Bendrat

S Rospert

W Buckel

Affiliations

Soon will be listed here.

Abstract

Membrane preparations of Fusobacterium nucleatum grown on glutamate contain glutaconyl-CoA decarboxylase at a high specific activity (13.8 nkat/mg protein). The enzyme was solubilized with 2% Triton X-100 in 0.5 M NaCl and purified 63-fold to a specific activity of 870 nkat/mg by affinity chromatography on monomeric avidin-Sepharose. The activity of the decarboxylase was strictly dependent on Na+ (Km = 3 mM) and was stimulated up to 3-fold by phospholipids. The glutaconyl-CoA decarboxylases from the gram-positive bacteria Acidaminococcus fermentans and Clostridium symbiosum have a lower apparent Km for Na+ (1 mM) and were not stimulated by phospholipids. In addition only the fusobacterial decarboxylase required sodium ion for stability and was inactivated by potassium ion. By incorporation of this purified enzyme into phospholipids an electrogenic sodium ion pump was reconstituted. The enzyme consists of four subunits, alpha (m = 65 kDa), beta (33 kDa), gamma (19 kDa), and delta (16 kDa) with the functions of a carboxy transferase (alpha), a carboxy lyase (beta and probably delta) and a biotin carrier (gamma). The subunits are very similar to those of the glutaconyl-CoA decarboxylases from the gram-positive bacteria. With an antiserum directed against the decarboxylase from A. fermentans the alpha- and the biotin containing subunits of the three decarboxylases and that from Peptostreptococcus asaccharolyticus could be detected on Western blots.

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References

Laemmli U . Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227(5259):680-5. DOI: 10.1038/227680a0. View

Forbush 3rd B . Assay of Na,K-ATPase in plasma membrane preparations: increasing the permeability of membrane vesicles using sodium dodecyl sulfate buffered with bovine serum albumin. Anal Biochem. 1983; 128(1):159-63. DOI: 10.1016/0003-2697(83)90356-1. View

Wifling K, Dimroth P . Isolation and characterization of oxaloacetate decarboxylase of Salmonella typhimurium, a sodium ion pump. Arch Microbiol. 1989; 152(6):584-8. DOI: 10.1007/BF00425491. View

Gravel R, Lam K, Mahuran D, Kronis A . Purification of human liver propionyl-CoA carboxylase by carbon tetrachloride extraction and monomeric avidin affinity chromatography. Arch Biochem Biophys. 1980; 201(2):669-73. DOI: 10.1016/0003-9861(80)90557-3. View

RACKER E, Violand B, ONeal S, Alfonzo M, Telford J . Reconstitution, a way of biochemical research; some new approaches to membrane-bound enzymes. Arch Biochem Biophys. 1979; 198(2):470-7. DOI: 10.1016/0003-9861(79)90521-6. View

Buckel W, Semmler R . Purification, characterisation and reconstitution of glutaconyl-CoA decarboxylase, a biotin-dependent sodium pump from anaerobic bacteria. Eur J Biochem. 1983; 136(2):427-34. DOI: 10.1111/j.1432-1033.1983.tb07760.x. View

Woese C . Bacterial evolution. Microbiol Rev. 1987; 51(2):221-71. PMC: 373105. DOI: 10.1128/mr.51.2.221-271.1987. View

Towbin H, Staehelin T, Gordon J . Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979; 76(9):4350-4. PMC: 411572. DOI: 10.1073/pnas.76.9.4350. View

Wohlfarth G, Buckel W . A sodium ion gradient as energy source for Peptostreptococcus asaccharolyticus. Arch Microbiol. 1985; 142(2):128-35. DOI: 10.1007/BF00447055. View

10.

Laussermair E, Schwarz E, Oesterhelt D, Reinke H, Beyreuther K, Dimroth P . The sodium ion translocating oxaloacetate decarboxylase of Klebsiella pneumoniae. Sequence of the integral membrane-bound subunits beta and gamma. J Biol Chem. 1989; 264(25):14710-5. View

11.

Buckel W, Semmler R . A biotin-dependent sodium pump: glutaconyl-CoA decarboxylase from Acidaminococcus fermentans. FEBS Lett. 1982; 148(1):35-8. DOI: 10.1016/0014-5793(82)81237-4. View

12.

Buckel W, Barker H . Two pathways of glutamate fermentation by anaerobic bacteria. J Bacteriol. 1974; 117(3):1248-60. PMC: 246608. DOI: 10.1128/jb.117.3.1248-1260.1974. View

13.

Buckel W, Liedtke H . The sodium pump glutaconyl-CoA decarboxylase from Acidaminococcus fermentans. Specific cleavage by n-alkanols. Eur J Biochem. 1986; 156(2):251-7. DOI: 10.1111/j.1432-1033.1986.tb09575.x. View

14.

Dzink J, Sheenan M, Socransky S . Proposal of three subspecies of Fusobacterium nucleatum Knorr 1922: Fusobacterium nucleatum subsp. nucleatum subsp. nov., comb. nov.; Fusobacterium nucleatum subsp. polymorphum subsp. nov., nom. rev., comb. nov.; and Fusobacterium nucleatum subsp..... Int J Syst Bacteriol. 1990; 40(1):74-8. DOI: 10.1099/00207713-40-1-74. View

15.

Van Veldhoven P, Mannaerts G . Inorganic and organic phosphate measurements in the nanomolar range. Anal Biochem. 1987; 161(1):45-8. DOI: 10.1016/0003-2697(87)90649-x. View

16.

Wang C, Smith R . Lowry determination of protein in the presence of Triton X-100. Anal Biochem. 1975; 63(2):414-7. DOI: 10.1016/0003-2697(75)90363-2. View

17.

Muller V, Blaut M, Gottschalk G . The transmembrane electrochemical gradient of Na+ as driving force for methanol oxidation in Methanosarcina barkeri. Eur J Biochem. 1988; 172(3):601-6. DOI: 10.1111/j.1432-1033.1988.tb13931.x. View

18.

Van den Eynde H, De Baere R, Shah H, Gharbia S, Fox G, Michalik J . 5S ribosomal ribonucleic acid sequences in Bacteroides and Fusobacterium: evolutionary relationships within these genera and among eubacteria in general. Int J Syst Bacteriol. 1989; 39(1):78-84. DOI: 10.1099/00207713-39-1-78. View

19.

Dimroth P . Sodium ion transport decarboxylases and other aspects of sodium ion cycling in bacteria. Microbiol Rev. 1987; 51(3):320-40. PMC: 373114. DOI: 10.1128/mr.51.3.320-340.1987. View

20.

Wessel D, Flugge U . A method for the quantitative recovery of protein in dilute solution in the presence of detergents and lipids. Anal Biochem. 1984; 138(1):141-3. DOI: 10.1016/0003-2697(84)90782-6. View