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α-Hemolysin: A Self-Assembling Protein Pore With Potential Applications In The Synthesis of New Materials

Published online by Cambridge University Press:  21 February 2011

Hagan Bayley ,
Musti Krishnasastry ,
Barbara Walker  and
John Kasianowicz
Hagan Bayley
Affiliation:
Worcester Foundation for Experimental Biology, 222 Maple Avenue, Shrewsbury, MA 01545
Musti Krishnasastry
Affiliation:
Worcester Foundation for Experimental Biology, 222 Maple Avenue, Shrewsbury, MA 01545
Barbara Walker
Affiliation:
Worcester Foundation for Experimental Biology, 222 Maple Avenue, Shrewsbury, MA 01545
John Kasianowicz
Affiliation:
National Institutes of Health, NIDDK-LBM, Bldg. 10-9B07, Bethesda, MD 20892
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Abstract

A selection of nanoscale membrane pores is being constructed by genetic manipulation of α-hemolysin (αHL), a 33.2 kDa polypeptide secreted by the bacterium Staphylococcus aureus, which can self-assemble into hexameric cylindrical channels -1 to 2 nm In Internal diameter. Ultimately, the new pores will be used to confer novel permeability properties upon materials such as thin films utilizing, for example, monolayer sheets of the hexamer. Recombinant αHL (r-αHL) has now been obtained in multimilligram amounts and purified to homogeneity after overexpression of the αHL gene in Escherichia coli. The properties of r-αHL are closely similar to those of αHL purified from S. aureus. Recent deletion mutagenesis experiments have given us new insight into the assembly mechanism of the pore. Three intermediates have been identified: a membrane-bound monomer; an oligomeric pore precursor; and the hexameric pore itself. Currently, point mutogenesis combined with chemical modification is being used to produce new pores of different internal diameter, with selectivity for the passage of molecules and Ions, and with gating properties (the ability to open and close in response to a physical stimulus, e.g. an electric field or light).

Type
Research Article
Information
MRS Online Proceedings Library (OPL) , Volume 255: Symposium Z – Hierarchically Structured Materials , 1991 , 201
Copyright
Copyright © Materials Research Society 1992

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