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Cloning, purification, and preliminary characterization by circular dichroism and NMR of a carboxyl‐terminal domain of the bacteriophage P22 scaffolding protein
Author(s) -
Parker Matthew H.,
Prevelige Peter E.,
Jablonsky Michael,
Casens Sherwood,
Sampson Laura,
Krish. Rama
Publication year - 1997
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1002/pro.5560060722
Subject(s) - circular dichroism , heteronuclear single quantum coherence spectroscopy , bacteriophage , protein secondary structure , scaffold protein , amino acid , filamentous bacteriophage , chemistry , scaffold , cloning (programming) , stereochemistry , crystallography , biophysics , biochemistry , nuclear magnetic resonance spectroscopy , biology , escherichia coli , medicine , signal transduction , computer science , biomedical engineering , gene , programming language
Assembly of double‐stranded DNA viruses and bacteriophages involves the polymerization of several hundred molecules of coat protein, directed by an internal scaffolding protein. A 163‐amino acid carboxyl‐terminal fragment of the 303‐amino acid bacteriophage P22 scaffolding protein was cloned, over‐expressed, and purified. This fragment is active in procapsid assembly reactions in vitro. The circular dichroism spectrum of the fragment, as well as the 1D‐NMR and 15 N‐ 1 H HSQC spectra of the uniformly‐labeled protein, indicate that stable secondary structure elements are present. Determination of the three dimensional packing of these elements into the folded scaffolding protein fragment is underway. Structure‐based drug design targeted at structural proteins required for viral assembly may have potential as a therapeutic strategy.