Mutagenesis of the cysteine-rich clip domain in the Drosophila patterning protease, Snake

Sufang Tian, Ellen K. LeMosy

Research output: Contribution to journalArticle

5 Scopus citations

Abstract

A common motif found in invertebrate serine proteases involved in immunity and development is the clip domain, proposed to regulate catalytic activity or protein-protein interactions within proteolytic cascades. Snake functions in a cascade that patterns the Drosophila embryo, and provides an accessible model for exploring the structural requirements for clip domain function. We tested Snake zymogens bearing charged-to-alanine mutations in the clip domain for their ability to rescue embryos lacking endogenous Snake and for their interactions by S2 cell co-transfection with upstream Gastrulation Defective and downstream Easter in the protease cascade. Of 13 single and multiple substitutions, one double mutant in a predicted protruding region exhibited a severe defect in embryonic rescue but showed only minimal defects in the co-transfection assay. We discuss implications of these and other results for potential biological roles of the Snake clip domain and for use of the in vitro assay in predicting protease behavior.

Original languageEnglish (US)
Pages (from-to)169-174
Number of pages6
JournalArchives of Biochemistry and Biophysics
Volume475
Issue number2
DOIs
StatePublished - Jul 15 2008

Keywords

  • Charged-to-alanine mutagenesis
  • Clip domain
  • Dorsoventral polarity
  • Drosophila
  • Serine protease

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology

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