| | The N-terminal domain of human holocarboxylase synthetase facilitates biotinylation via direct interaction with the substrate proteinEdited by Gianni Cesareni Received 10 November 2009; received in revised form 21 December 2009; accepted 26 December 2009. published online 18 January 2010. Abstract Human holocarboxylase synthetase shows a high degree of sequence homology in the catalytic domain with bacterial biotin ligases such as Escherichia coli BirA, but differs in the length and sequence of the N-terminus. Despite several studies having been undertaken on the N-terminal region of hHCS, the role of this region remains unclear. We determined the structure of the N-terminal domain of hHCS by limited proteolysis and showed that this domain has a crucial effect on the enzymatic activity. The domain interacts not only with biotin acceptor protein, but also with the catalytic domain of hHCS, as shown by nuclear magnetic resonance (NMR) experiments. We propose that the N-terminal domain of hHCS recognizes the charged region of biotin acceptor protein, distinctly from the recognition by the catalytic domain. Division of Magnetic Resonance Research, Korea Basic Science Institute, 804-1 Yangcheong-Ri, Ochang-Eup, Cheongwon-Gun, Chungbuk 363-883, Republic of Korea Bio-Analytical Science Program, University of Science and Technology, Daejeon 350-333, Republic of Korea  Corresponding authors. Address: Division of Magnetic Resonance Research, Korea Basic Science Institute, 804-1 Yangcheong-Ri, Ochang-Eup, Cheongwon-Gun, Chungbuk 363-883, Republic of Korea. Fax: +82 43 240 5059.
PII: S0014-5793(10)00047-5 doi:10.1016/j.febslet.2009.12.059 © 2010 Federation of European Biochemical Societies | |
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ABSTRACT
