Preliminary X-ray crystallographic analysis of biotin carboxylase isolated from Escherichia coli.
Acetyl CoA carboxylase catalyzes the first committed step in the biosynthesis of long chain fatty acids. In Escherichia coli, the enzyme consists of three subunits that are isolated separately and display distinct functional properties. Here we report the crystallization and preliminary X-ray analysis of one of these components, namely biotin carboxylase. The crystals are grown by microdialysis against 10 mM potassium phosphate (pH 7.0), 1 mM EDTA, 2 mM DTT and 1 mM NaN3 at 4 degrees C. They belong to the space group P2(1)2(1)2(1) with unit cell dimensions of a = 61.9 A, b = 96.1 A and c = 180.6 A and contain one dimer per asymmetric unit. The crystals diffract to a nominal resolution of 2.2 A. From a mechanistic standpoint, biotin carboxylase is especially interesting in that it is the smallest protein within its class and is one of only two carboxylases that can utilize free biotin as a substrate.
Waldrop
G
G
Institute for Enzyme Research, Graduate School and the Department of Biochemistry, University of Wisconsin, Madison 53705.
Holden
H M
HM
Rayment
I
I
D000103:Acetyl-CoA Carboxylase / Q000737:chemistry / Q000302:isolation & purification; D019731:Carbon-Nitrogen Ligases; D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods; D004926:Escherichia coli / Q000201:enzymology*; D008025:Ligases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D046911:Macromolecular Substances; D011487:Protein Conformation
D046911:Macromolecular Substances; D008025:Ligases; D019731:Carbon-Nitrogen Ligases; C019791:biotin carboxylase; D000103:Acetyl-CoA Carboxylase