DOI: 10.1111/j.1432-1033.1995.828_3.x. Purification of Active E1α2β2 of Pseudomonas Putida Branched‐Chain‐Oxoacid Dehydrogenase

Purification of Active E1α2β2 of Pseudomonas Putida Branched‐Chain‐Oxoacid Dehydrogenase

10.1111/j.1432-1033.1995.828_3.x

Crossref journal-article

Wiley

European Journal of Biochemistry (311)

Abstract

Active E1 component of Pseudomonas putida branched‐chain‐oxoacid dehydrogenase was purified from P. putida strains carrying pJRS84 which contains bkdR (encoding the transcriptional activator) and bkdA1 and bkdA2 (encoding the α and β) subunits). Expression was inducible, however, 45– 39– and 37‐kDa proteins were produced instead of the expected 45‐kDa and 37‐kDa proteins. The 45‐kDa protein was identified as E1α and the 37‐kDa and 39‐kDa proteins were identified as separate translational products of bkdA2 by their N‐terminal sequences. The N‐terminal amino acid of the 39‐kDa protein was leucine instead of methionine. The 45–, 39– and 37‐kDa proteins were also produced in wild‐type P. putida. Translation of bkdA1 and bkdA2 from an Escherichia coli expression plasmid produced only 45‐kDa and 39‐kDa proteins, with N‐terminal methionine on the 39‐kDa protein. The insertion of guanine residues 5′ to the first ATG of bkdA2 did not affect expression of E1β in P. putida including the N‐terminal leucine which appears to eliminate the possibility of ribosome jumping. The Z‐average molecular mass of the El component was determined by sedimentation equilibrium to be 172±9 kDa compared to a calculated value of 166 kDa for the heterotetramer and a Stokes radius of 5.1 nm. E1α Ser313, which is homologous to the phosphorylated residue of rat liver E1α, was converted to alanine resulting in about a twofold increase in Km, but no change in Kcat. S315A and S319A mutations had no effect on Km or Kcat, indicating that these residues do not play a major part in catalysis of E1α2β2.

Bibliography

Hester, K., Luo, J., Burns, G., Braswell, E. H., & Sokatch, J. R. (1995). Purification of Active E1Î±2Î²2 of Pseudomonas Putida BranchedâChainâOxoacid Dehydrogenase. European Journal of Biochemistry, 233(3), 828â836. Portico.

Authors 5

Kathryn Hester (first)
Jinhe Luo (additional)
Gayle Burns (additional)
Emory H. Braswell (additional)
John R. Sokatch (additional)

References 58 Referenced 12

{'key': 'e_1_2_3_2_2', 'first-page': '671', 'volume-title': 'The metabolic basis of inherited diesease', 'author': 'Danner D. J.', 'year': '1989'} / The metabolic basis of inherited diesease by Danner D. J. (1989)
{'key': 'e_1_2_3_3_2', 'first-page': '869', 'volume-title': 'The metabolic basis of inherited diesease', 'author': 'Robinson B. H.', 'year': '1989'} / The metabolic basis of inherited diesease by Robinson B. H. (1989)
10.1016/0959-440X(92)90114-M
{'key': 'e_1_2_3_5_2', 'first-page': '121', 'volume-title': 'Chemistry and biochemistry of flavoenzymes', 'author': 'Williams C. H. J.', 'year': '1992'} / Chemistry and biochemistry of flavoenzymes by Williams C. H. J. (1992)
10.1128/jb.173.10.3109-3116.1991 / J. Bacteriol. / Cloning and sequence analysis of the LPD‐glc structural gene of Pseudomonas putida by Palmer J. A. (1991)
10.1016/0003-9861(84)90036-5
10.1111/j.1432-1033.1989.tb14521.x
10.1128/jb.148.2.639-646.1981 / J. Bacteriol. / Isolation of a specific lipoamide dehydrogenase for a branched‐chain keto acid dehydrogenase from Pseudomonas putida by Sokatch J. R. (1981)
10.1016/0022-2836(89)90486-5
10.1016/0022-2836(91)90367-F
10.1002/prot.340130406
10.1006/jmbi.1993.1236
10.1126/science.1549782
10.1111/j.1432-1033.1983.tb07441.x
10.1111/j.1432-1033.1985.tb09160.x
{'key': 'e_1_2_3_17_2', 'first-page': '393', 'article-title': 'The pyruvate dehydrogenase complex of Pseudomonas aeruginosa PAO. Purification, properties and characterization of mutants', 'volume': '120', 'author': 'Jeyaseelan K.', 'year': '1980', 'journal-title': 'J. Gen. Microbiol.'} / J. Gen. Microbiol. / The pyruvate dehydrogenase complex of Pseudomonas aeruginosa PAO. Purification, properties and characterization of mutants by Jeyaseelan K. (1980)
10.1111/j.1432-1033.1990.tb19128.x
10.1111/j.1432-1033.1988.tb14283.x
10.1021/bi00605a017
10.1111/j.1432-1033.1984.tb08597.x
10.1042/bj2040353
10.1016/0003-9861(86)90108-6
10.1128/JB.148.2.647-652.1981
10.1016/0003-9861(72)90152-X
10.1016/S0021-9258(18)42045-5 / J. Biol. Chem. / Expression and assembly of a functional E1 component (α2β2) of mammalian branched‐chain α‐ketoacid dehydrogenase complex in Escherichia coli by Davie J. R. (1992)
10.1016/S0021-9258(17)34071-1 / J. Biol. Chem / Expression in Escherichia coli of genes encoding the E1α and E1β subunits of the pyruvate dehydrogenase complex of Bacillus stearorhermophilus and assembly of a functional El component (α2β2) in vitro by Lessard I. A. D. (1994)
10.1111/j.1432-1033.1988.tb14264.x
10.1128/jb.175.13.3934-3940.1993
10.1128/jb.177.3.636-641.1995
10.1128/jb.110.3.1073-1081.1972 / J. Bacteriol. / Regulation of valine catabolism in Pseudomonas putida by Marshall V. P. (1972)
10.1016/0378-1119(93)90476-J
10.1016/S0021-9258(18)42252-1 / J. Biol. Chem. / Characterization of the mmsAB operon of Pseudonomas aeruginosa PAO encoding methylmalonate‐semialdehyde dehydrogenase and 3‐hydroxyisobutyrate dehydrogenase by Steele M. I. (1992)
{'key': 'e_1_2_3_34_2', 'volume-title': 'Molecular cloning: a laboratory manual', 'author': 'Sambrook J.', 'year': '1989'} / Molecular cloning: a laboratory manual by Sambrook J. (1989)
10.1128/jb.172.10.5655-5663.1990 / J. Bucteriol. / Transcriptional analysis of the promoter region of the Pseudomonus putida branched‐chain keto acid dehydrogenase operon by Madhusudhan K. T. (1990)
10.1016/0378-1119(82)90015-4
10.1016/0378-1119(81)90080-9
10.1016/0147-619X(91)90056-3 / Plasmid / A fully modular vector system for the optimization of gene expression in Escherichia coli by Belev T. N. (1992)
{'key': 'e_1_2_3_39_2', 'first-page': '177', 'volume-title': 'PCR protocols a guide to methods and applications', 'author': 'Higuchi R.', 'year': '1990'} / PCR protocols a guide to methods and applications by Higuchi R. (1990)
10.1016/0042-6822(55)90016-7 / Virology / Transduction of linked genetic characters of the host by bacteriophage P1 by Lennox E. S. (1955)
10.1128/jb.169.4.1619-1625.1987
10.1016/0003-2697(76)90527-3
10.1016/0014-5793(84)80259-8
10.1016/0003-2697(70)90103-X
10.1016/S0006-3495(81)84753-4
{'key': 'e_1_2_3_46_2', 'first-page': '370', 'volume-title': 'Proteins, amino acids and peptides as ions and dipolar ions', 'author': 'Cohn E. J.', 'year': '1943'} / Proteins, amino acids and peptides as ions and dipolar ions by Cohn E. J. (1943)
10.1016/0003-2697(92)90316-Y
10.1016/B978-0-08-092461-8.50010-1
10.1042/bj2330737
10.1016/S0076-6879(88)66047-2 / Methods Enzymol. / Cloning of genes for branched‐chain keto acid dehydrogenase in Pseudomonas putida by Sykes P. J. (1988)
10.1016/0378-1119(83)90197-X
10.1016/S0021-9258(17)32349-9 / J. Biol. Chem. / Site‐directed mutagenesis of phosphorylation sites of the branched chain α‐ketoacid dehydrogenase complex by Zhao Y. (1994)
10.1126/science.2830666
10.1016/0092-8674(90)90245-A
10.1016/S0021-9258(19)45399-4 / J. Biol. Chem. / Isolation of rabbit liver branched chain α‐ketoacid dehydrogenase and regulation by phosphorylation by Paxton R. (1982)
10.1042/bj1920155
10.1146/annurev.bi.57.070188.001215
10.1016/S0021-9258(18)48161-6
10.1016/0076-6879(90)83007-V

Dates

Type	When
Created	20 years, 11 months ago (Sept. 7, 2004, 8:20 a.m.)
Deposited	1 year, 10 months ago (Oct. 7, 2023, 4:11 p.m.)
Indexed	1 year, 7 months ago (Jan. 9, 2024, 2:23 p.m.)
Issued	29 years, 9 months ago (Nov. 1, 1995)
Published	29 years, 9 months ago (Nov. 1, 1995)
Published Online	20 years, 11 months ago (Sept. 7, 2004)
Published Print	29 years, 9 months ago (Nov. 1, 1995)

Funders 0

None

BibTeX

@article{Hester_1995, title={Purification of Active E1α2β2 of Pseudomonas Putida Branched‐Chain‐Oxoacid Dehydrogenase}, volume={233}, ISSN={1432-1033}, url={http://dx.doi.org/10.1111/j.1432-1033.1995.828_3.x}, DOI={10.1111/j.1432-1033.1995.828_3.x}, number={3}, journal={European Journal of Biochemistry}, publisher={Wiley}, author={Hester, Kathryn and Luo, Jinhe and Burns, Gayle and Braswell, Emory H. and Sokatch, John R.}, year={1995}, month=nov, pages={828–836} }

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