A single point mutation in a Group I WW domain shifts its specificity to that of Group II WW domains

Xavier Espanel, Marius Sudol

Research output: Contribution to journalArticlepeer-review

54 Scopus citations

Abstract

WW domains can be divided into three groups based on their binding specificity. By random mutagenesis, we switched the specificity of the Yes- associated protein (YAP) WW1 domain, a Group I WW domain, to that of the FE65 WW domain, which belongs to Group II. We showed that a single mutation, leucine 190 (βB5) to tryptophan, is required to switch from Group I to Group II. Although this single substitution in YAP WW1 domain is sufficient to precipitate the two protein isoforms of Mena, an in vivo ligand of FE65, we showed that an additional substitution, histidine 192 (βB7) to glycine, significantly increased the ability of YAP to mimic FE65. This double mutant (L190W/H192G) precipitates eight of the nine protein bands that FE65 pulls down from rat brain protein lysates. Based on both our data and a sequence comparison between Group I and Group II WW domains, we propose that a block of three consecutive aromatic amino acids within the second β-sheet of the domain is required, but not always sufficient, for a WW domain to belong to Group II. These data deepen our understanding of WW domain binding specificity and provide a basis for the rational design of modified WW domains with potential therapeutic applications.

Original languageEnglish
Pages (from-to)17284-17289
Number of pages6
JournalJournal of Biological Chemistry
Volume274
Issue number24
DOIs
StatePublished - 11 Jun 1999

Fingerprint

Dive into the research topics of 'A single point mutation in a Group I WW domain shifts its specificity to that of Group II WW domains'. Together they form a unique fingerprint.

Cite this