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Proteins. 2016 Oct;84(10):1422-30. doi: 10.1002/prot.25087. Epub 2016 Jul 5.

Characterization of the structure and catalytic activity of Legionella pneumophila VipF.

Author information

1
Department of Chemistry and Biochemistry, James Madison University, Harrisonburg, Virginia, 22807.
2
Department of Chemistry and Biochemistry, James Madison University, Harrisonburg, Virginia, 22807. berndsce@jmu.edu.

Abstract

The pathogenic bacteria Legionella pneumophila is known to cause Legionnaires' Disease, a severe pneumonia that can be fatal to immunocompromised individuals and the elderly. Shohdy et al. identified the L. pneumophila vacuole sorting inhibitory protein VipF as a putative N-acetyltransferase based on sequence homology. We have characterized the basic structural and functional properties of VipF to confirm this original functional assignment. Sequence conservation analysis indicates two putative CoA-binding regions within VipF. Homology modeling and small angle X-ray scattering suggest a monomeric, dual-domain structure joined by a flexible linker. Each domain contains the characteristic beta-splay motif found in many acetyltransferases, suggesting that VipF may contain two active sites. Docking experiments suggest reasonable acetyl-CoA binding locations within each beta-splay motif. Broad substrate screening indicated that VipF is capable of acetylating chloramphenicol and both domains are catalytically active. Given that chloramphenicol is not known to be N-acetylated, this is a surprising finding suggesting that VipF is capable of O-acetyltransferase activity. Proteins 2016; 84:1422-1430.

KEYWORDS:

acetyltransferase; chloramphenicol; legionella; molecular modeling; small angle X-ray scattering

PMID:
27315603
DOI:
10.1002/prot.25087
[Indexed for MEDLINE]

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