TY - JOUR
T1 - Biotin protein ligase from Candida albicans: Expression, purification and development of a novel assay
AU - Pendini, Nicole R
AU - Bailey, Lisa M
AU - Booker, Grant William
AU - Wilce, Matthew Charles James
AU - Wallace, John Campbell
AU - Polyak, Steven W
PY - 2008
Y1 - 2008
N2 - Biotin protein ligase (BPL) is an essential enzyme responsible for the activation of biotin-dependent enzymes through the covalent attachment of biotin. In yeast, disruption of BPL affects important metabolic pathways such as fatty acid biosynthesis and gluconeogenesis. This makes BPL an attractive drug target for new antifungal agents. Here we report the cloning, recombinant expression and purification of BPL from the fungal pathogen Candida albicans. The biotin domains of acetyl CoA carboxylase and pyruvate carboxylase were also cloned and characterised as substrates for BPL. A novel assay was established thereby allowing examination of the enzyme s properties. These findings will facilitate future structural studies as well as screening efforts to identify potential inhibitors.
AB - Biotin protein ligase (BPL) is an essential enzyme responsible for the activation of biotin-dependent enzymes through the covalent attachment of biotin. In yeast, disruption of BPL affects important metabolic pathways such as fatty acid biosynthesis and gluconeogenesis. This makes BPL an attractive drug target for new antifungal agents. Here we report the cloning, recombinant expression and purification of BPL from the fungal pathogen Candida albicans. The biotin domains of acetyl CoA carboxylase and pyruvate carboxylase were also cloned and characterised as substrates for BPL. A novel assay was established thereby allowing examination of the enzyme s properties. These findings will facilitate future structural studies as well as screening efforts to identify potential inhibitors.
UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18809372
M3 - Article
SN - 0003-9861
VL - 479
SP - 163
EP - 169
JO - Archives of Biochemistry and Biophysics
JF - Archives of Biochemistry and Biophysics
IS - 2
ER -