Preliminary crystallographic analysis of the BPL from Pyrococcus horikoshii OT3

What is it about?

Biotin±protein ligase is an enzyme that catalyzes the ATP-dependent biotinylation of a specific lysine residue in acetyl-CoA carboxylase. The biotin±protein ligase from Pyrococcus horikoshii OT3 has been cloned, overexpressed and puri®ed. Crystallization was performed by the microbatch method or the vapour-diffusion method using PEG 2000 as a precipitant at 295 K. X-ray diffraction data have been collected to 1.6 A resolution from a native crystal and to 1.55 A resolution from a selenomethionine-derivative crystal for multiple anomalous dispersion phasing using synchrotron radiation at 100 K

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Why is it important?

Biotin±protein ligase (BPL) mediates the biotinylation of acetyl-CoA carboxylase via the covalent attachment of biotin to a specific lysine residue of the biotin carboxyl carrier protein (BCCP), which is a core subunit of the carboxylase. This is a two-step reaction: adenylated biotin is first synthesized from the substrates biotin and ATP and the biotin moiety is then transferred to the specific lysine residue of BCCP. The reaction is highly conserved in all organisms, which is reinforced by the fact that BPLs from different species are able to recognize and correctly biotinylate carboxylases from widely divergent sources.

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