Heteronuclear NMR studies of the specificity of the post‐translational modification of biotinyl domains by biotinyl protein ligase

FEBS Letters

Howard, Mark J.¹  Broadhurst, R.William¹  Perham, Richard N.¹  Reche, Pedro A.¹ 

[1] Cambridge Centre for Molecular Recognition, Department of Biochemistry, University of Cambridge, 80 Tennis Court Road, Cambridge CB2 1GA, UK

关键词: Biotin;    Biotinyl protein ligase;    Protein domain;    Molecular recognition;    NMR spectroscopy;    BCCP;    biotin carboxyl carrier protein;    BPL;    biotinyl protein ligase;    GCSH;    the H protein of the glycine decarboxylase system;    HMQC;    heteronuclear multiple quantum coherence;    LPL;    lipoyl protein ligase;    NMR;    nuclear magnetic resonance;   

DOI  :  10.1016/S0014-5793(00)01829-9

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The lipoyl domains of 2-oxo acid dehydrogenase multienzyme complexes and the biotinyl domains of biotin-dependent enzymes have homologous structures, but the target lysine residue in each domain is correctly selected for post-translational modification by lipoyl protein ligase and biotinyl protein ligase, respectively. We have applied two-dimensional heteronuclear NMR spectroscopy to investigate the interaction between the apo form of the biotinyl domain of the biotin carboxyl carrier protein of acetyl-CoA carboxylase and the biotinyl protein ligase (BPL) from Escherichia coli. Heteronuclear multiple quantum coherence NMR spectra of the ¹⁵N-labelled biotinyl domain were recorded in the presence and absence of the ligase and backbone amide ¹H and ¹⁵N chemical shifts were evaluated. Small, but significant, changes in chemical shift were found in two regions, including the tight β-turn that houses the lysine residue targetted for biotinylation, and the β-strand 2 and the loop that precedes it in the domain. When compared with the three-dimensional structure, sequence alignments of other biotinyl and lipoyl domains, and mutagenesis data, these results give a clear indication of how the biotinyl domain is both recognised by BPL and distinguished from the structurally related lipoyl domain to ensure correct post-translational modification.

【 授权许可】

Unknown