Document Detail

A novel approach for over-expression, characterization, and isotopic enrichment of a homogeneous species of acyl carrier protein from Plasmodium falciparum.
MedLine Citation:
PMID:  15823545     Owner:  NLM     Status:  MEDLINE    
Acyl carrier protein (ACP) plays a central role in fatty acid biosynthesis by transferring the acyl groups from one enzyme to another for the completion of the fatty acid synthesis cycle. Holo-ACP is the obligatory substrate for the synthesis of acyl-ACPs which act as the carrier and donor for various metabolic reactions. Despite its interactions with numerous proteins in the cell, its mode of interaction is poorly understood. Here, we report the over-expression of PfACP in minimal medium solely in its holo form and in high yield. Expression in minimal media provides a means to isotopically label PfACP for high resolution multi-nuclear and multi-dimensional NMR studies. Indeed, the proton-nitrogen correlated NMR spectrum exhibits very high chemical shift dispersion and resolution. We also show that holo-PfACP thus expressed is amenable to acylation reactions using Escherichia coli acyl-ACP synthetase as well as by standard chemical methods.
Shailendra Kumar Sharma; Rahul Modak; Shilpi Sharma; Alok Kumar Sharma; Siddhartha P Sarma; Avadhesha Surolia; Namita Surolia
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Biochemical and biophysical research communications     Volume:  330     ISSN:  0006-291X     ISO Abbreviation:  Biochem. Biophys. Res. Commun.     Publication Date:  2005 May 
Date Detail:
Created Date:  2005-04-12     Completed Date:  2005-07-05     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0372516     Medline TA:  Biochem Biophys Res Commun     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1019-26     Citation Subset:  IM    
Molecular Biophysics Unit, Indian Institute of Science, Bangalore 560 012, India.
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MeSH Terms
Acyl Carrier Protein / biosynthesis*,  chemistry,  genetics
Carbon Isotopes
Carbon-Sulfur Ligases / biosynthesis
Circular Dichroism
Escherichia coli / genetics,  metabolism
Histidine / chemistry
Isotope Labeling
Nitrogen Isotopes
Nuclear Magnetic Resonance, Biomolecular
Plasmodium falciparum / metabolism*
Protein Folding
Recombinant Proteins / biosynthesis,  genetics
Reg. No./Substance:
0/Acyl Carrier Protein; 0/Carbon Isotopes; 0/Nitrogen Isotopes; 0/Recombinant Proteins; 71-00-1/Histidine; EC 6.2.-/Carbon-Sulfur Ligases; EC ligase

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