PMID: 9445386Mar 7, 1998Paper

Selectivity of post-translational modification in biotinylated proteins: the carboxy carrier protein of the acetyl-CoA carboxylase of Escherichia coli

The Biochemical Journal
P A RecheR N Perham

Abstract

Biotin-dependent enzymes contain a biotinyl-lysine residue in a conserved sequence motif, MKM, located in a surface hairpin turn in one of the two beta-sheets that make up the domain. A sub-gene encoding the 82-residue C-terminal biotinyl domain from the biotin carboxy carrier protein of acetyl-CoA carboxylase from Escherichia coli as a fusion protein with glutathione S-transferase was created and over-expressed in E. coli. The biotinyl domain was readily released by cleavage with thrombin. Five mutant domains were created in which the conserved MKM motif was systematically replaced: by MAK and KAM, in which the target lysine is moved one place; by KKM and MKK, in which a second potential site for biotinylation is introduced; and by DKA, the motif found in the correspondingly conserved site of lipoylation in the structurally related lipoyl domains of 2-oxo acid dehydrogenase multienzyme complexes. No biotinylation of the MAK or KAM mutants was observed in vivo or by purified biotinyl protein ligase in vitro; in the KKM and MKK mutants, only one lysine residue, presumed to be that in its native position in the hairpin turn, was found to be biotinylated in vivo and in vitro. The DKA mutant was not biotinylated in vivo, but was pa...Continue Reading

Citations

Apr 30, 2008·Biochimica Et Biophysica Acta·Nicole R PendiniSteven W Polyak
Oct 2, 2004·Protein Science : a Publication of the Protein Society·Eunjoo Choi-RheeJohn E Cronan
May 30, 2008·PloS One·Sudha PurushothamanAvadhesha Surolia
Nov 21, 2001·Protein Science : a Publication of the Protein Society·L H WeaverB W Matthews
Nov 21, 2001·Protein Science : a Publication of the Protein Society·A Chapman-SmithJ C Wallace
Mar 8, 2017·Trends in Biochemical Sciences·Louise M SternickiSteven W Polyak
Nov 2, 2018·Biochemical Society Transactions·Dorothy Beckett
Nov 3, 2011·Virology Journal·Larisa Smelyanski, Jonathan M Gershoni
Feb 17, 2006·FEBS Letters·Yan-Qiu LiYutaka Kawarabayasi
Sep 3, 1999·The Journal of Biological Chemistry·V GueguenJ Bourguignon
Jun 13, 2009·The Journal of Biological Chemistry·Quin H Christensen, John E Cronan
Mar 29, 2008·The Journal of Biological Chemistry·Bagautdin BagautdinovNaoki Kunishima
Mar 4, 1999·The Journal of Nutrition·A Chapman-Smith, J E Cronan
Sep 2, 1999·Trends in Biochemical Sciences·A Chapman-Smith, J E Cronan
Aug 2, 2002·Biochemical and Biophysical Research Communications·Sarawut JitrapakdeeJohn C Wallace

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