Chemical-genetic inhibition of sensitized mutant unconventional myosins.

Ryan L. Karcher, D. William Provance, Peter Barr-Gillespie, John A. Mercer

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

The construction of a sensitized mutant unconventional myosin is an excellent method for determining the function of the individual myosin against a background of related myosins with partially overlapping functions. In this chapter, we outline the steps involved in sensitizing myosin by mutation and screening them against panels of nucleotide analogs, including transfection, microinjection, and actin co-sedimentation in vitro. We also describe conditions and considerations involved in designing functional experiments after a mutant and cognate analog have been identified. The powerful strategy of chemical genetics, when correctly applied to unconventional myosins, enables both the specific and selectable inhibition of the target motor with outstanding internal controls.

Original languageEnglish (US)
Pages (from-to)231-240
Number of pages10
JournalMethods in molecular biology (Clifton, N.J.)
Volume392
StatePublished - 2007
Externally publishedYes

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Myosins
Microinjections
Transfection
Actins
Nucleotides
Mutation

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Chemical-genetic inhibition of sensitized mutant unconventional myosins. / Karcher, Ryan L.; Provance, D. William; Barr-Gillespie, Peter; Mercer, John A.

In: Methods in molecular biology (Clifton, N.J.), Vol. 392, 2007, p. 231-240.

Research output: Contribution to journalArticle

Karcher, Ryan L. ; Provance, D. William ; Barr-Gillespie, Peter ; Mercer, John A. / Chemical-genetic inhibition of sensitized mutant unconventional myosins. In: Methods in molecular biology (Clifton, N.J.). 2007 ; Vol. 392. pp. 231-240.
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