Journal
PROTEIN EXPRESSION AND PURIFICATION
Volume 36, Issue 1, Pages 100-105Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.pep.2004.03.007
Keywords
N-formylmethionine; peptide deformylase; N-terminal processing; rubredoxin
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Funding
- NIGMS NIH HHS [GM 64736] Funding Source: Medline
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Deformylation of the initiator N-formylmethionine does not always proceed to completion for proteins overexpressed in Escherichia coli. To overcome this limitation, the def gene encoding the Escherichia coli peptide deformylase was cloned into the plysS plasmid under the tetracycline (Tc) promoter control. The efficiency of this constitutive level of peptide deformylase expression was demonstrated for the case of the rubredoxins from both mesophilic and hyperthermophilic organisms which normally retain a majority of their N-formyl terminal form. Indicating the potential structural/functional significance of residual formylation, the presence of a highly solvent exposed N-formyl group in rubredoxin is discernable in the amide NMR chemical shifts for the active site metal-coordinating cysteines more than 21 Angstrom away. (C) 2004 Elsevier Inc. All rights reserved.
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