Please use this identifier to cite or link to this item: http://hdl.handle.net/2440/11303
Citations
Scopus Web of Science® Altmetric
?
?
Type: Journal article
Title: The biotin domain peptide from the biotin carboxyl carrier protein of Escherichia coli acetyl-CoA carboxylase causes a marked increase in the catalytic efficiency of biotin carboxylase and carboxyltransferase relative to free biotin
Author: Blanchard, C.
Chapman-Smith, A.
Wallace, J.
Waldrop, G.
Citation: Journal of Biological Chemistry, 1999; 274(45):31767-31769
Publisher: AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
Issue Date: 1999
ISSN: 0021-9258
1083-351X
Statement of
Responsibility: 
Carol Z. Blanchard, Anne Chapman-Smith, John C. Wallace and Grover L. Waldrop
Abstract: Acetyl-CoA carboxylase catalyzes the first committed step in the biosynthesis of long-chain fatty acids. The Escherichia coli form of the enzyme consists of a biotin carboxylase activity, a biotin carboxyl carrier protein, and a carboxyltransferase activity. The C-terminal 87 amino acids of the biotin carboxyl carrier protein (BCCP87) form a domain that can be independently expressed, biotinylated, and purified (Chapman-Smith, A., Turner, D. L., Cronan, J. E., Morris, T. W., and Wallace, J. C. (1994) Biochem. J. 302, 881-887). The ability of the biotinylated form of this 87-residue protein (holoBCCP87) to act as a substrate for biotin carboxylase and carboxyltransferase was assessed and compared with the results with free biotin. In the case of biotin carboxylase holoBCCP87 was an excellent substrate with a K(m) of 0.16 +/- 0.05 mM and V(max) of 1000.8 +/- 182.0 min(-1). The V/K or catalytic efficiency of biotin carboxylase with holoBCCP87 as substrate was 8000-fold greater than with biotin as substrate. Stimulation of the ATP synthesis reaction of biotin carboxylase where carbamyl phosphate reacted with ADP by holoBCCP87 was 5-fold greater than with an equivalent amount of biotin. The interaction of holoBCCP87 with carboxyltransferase was characterized in the reverse direction where malonyl-CoA reacted with holoBCCP87 to form acetyl-CoA and carboxyholoBCCP87. The K(m) for holoBCCP87 was 0.45 +/- 0.07 mM while the V(max) was 2031.8 +/- 231.0 min(-1). The V/K or catalytic efficiency of carboxyltransferase with holoBCCP87 as substrate is 2000-fold greater than with biotin as substrate.
Keywords: Acetyl-coa Carboxylase, Metabolism, Biotin, Metabolism, Carbon-nitrogen Ligases, Metabolism, Carboxyl And Carbamoyl Transferases, Metabolism, Carrier Proteins, Metabolism, Peptide Fragments, Metabolism
Rights: © The American Society for Biochemistry and Molecular Biology, Inc.
RMID: 0030004401
DOI: 10.1074/jbc.274.45.31767
Appears in Collections:Biochemistry publications

Files in This Item:
There are no files associated with this item.


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.