PMID:12515551
| Citation |
Worsham, LM, Earls, L, Jolly, C, Langston, KG, Trent, MS and Ernst-Fonberg, ML (2003) Amino acid residues of Escherichia coli acyl carrier protein involved in heterologous protein interactions. Biochemistry 42:167-76 |
|---|---|
| Abstract |
Acyl carrier protein (ACP) is a small, highly conserved protein with an essential role in a myriad of reactions throughout lipid metabolism in plants and bacteria where it interacts with a remarkable diversity of proteins. The nature of the proper recognition and precise alignment between the protein moieties of ACP and its many interactive proteins is not understood. Residues conserved among ACPs from numerous plants and bacteria were considered as possibly being crucial to ACP's function, including protein-protein interaction, and a method of identifying amino acid residue clusters of high hydrophobicity on ACP's surface was used to estimate residues possibly involved in specific ACP-protein interactions. On the basis of this information, single-site mutation analysis of multiple residues, one at a time, of ACP was used to probe the identities of potential contact residues of ACPSH or acyl-ACP involved in specific interactions with selected enzymes. The roles of particular ACP residues were more precisely defined by site-directed fluorescence analyses of various myristoyl-mutant-ACPs upon specific interaction with the Escherichia coli hemolysin-activating acyltransferase, HlyC. This was done by selectively labeling each mutated site, one at a time, with an environmentally sensitive fluoroprobe and observing its fluorescence behavior in the absence and presence of HlyC. Consequently, a picture of the portion of ACP involved in selected macromolecular interaction has emerged. |
| Links |
PubMed Online version:10.1021/bi0261950 |
| Keywords |
Acyl Carrier Protein/chemistry; Acyl Carrier Protein/genetics; Acyl Carrier Protein/metabolism; Acylation; Amino Acid Sequence; Amino Acid Substitution/genetics; Amino Acids/chemistry; Amino Acids/genetics; Amino Acids/metabolism; Anilino Naphthalenesulfonates/metabolism; Escherichia coli Proteins/chemistry; Escherichia coli Proteins/genetics; Escherichia coli Proteins/metabolism; Fatty Acid Synthetase Complex/metabolism; Fluorescence Polarization; Fluorescent Dyes/metabolism; Hemolysin Proteins/metabolism; Molecular Sequence Data; Mutagenesis, Site-Directed; Myristic Acid/metabolism; Spectrometry, Fluorescence; Substrate Specificity/genetics; Sulfhydryl Compounds/chemistry; Sulfhydryl Reagents/metabolism |
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