Structural basis for small G protein effector interaction of Ras-related protein 1 (Rap1) and adaptor protein Krev interaction trapped 1 (KRIT1)

J Biol Chem. 2012 Jun 22;287(26):22317-27. doi: 10.1074/jbc.M112.361295. Epub 2012 May 10.

Abstract

Cerebral cavernous malformations (CCMs) affect 0.1-0.5% of the population resulting in leaky vasculature and severe neurological defects. KRIT1 (Krev interaction trapped-1) mutations associate with ∼40% of familial CCMs. KRIT1 is an effector of Ras-related protein 1 (Rap1) GTPase. Rap1 relocalizes KRIT1 from microtubules to cell membranes to impact integrin activation, potentially important for CCM pathology. We report the 1.95 Å co-crystal structure of KRIT1 FERM domain in complex with Rap1. Rap1-KRIT1 interaction encompasses an extended surface, including Rap1 Switch I and II and KRIT1 FERM F1 and F2 lobes. Rap1 binds KRIT1-F1 lobe using a GTPase-ubiquitin-like fold interaction but binds KRIT1-F2 lobe by a novel interaction. Point mutagenesis confirms the interaction. High similarity between KRIT1-F2/F3 and talin is revealed. Additionally, the mechanism for FERM domains acting as GTPase effectors is suggested. Finally, structure-based alignment of each lobe suggests classification of FERM domains as ERM-like and TMFK-like (talin-myosin-FAK-KRIT-like) and that FERM lobes resemble domain "modules."

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Crystallography, X-Ray / methods
  • GTP Phosphohydrolases / metabolism
  • Gene Expression Regulation*
  • Hemangioma, Cavernous, Central Nervous System / metabolism
  • Humans
  • Integrins / metabolism
  • KRIT1 Protein
  • Microtubule-Associated Proteins / metabolism*
  • Models, Biological
  • Models, Molecular
  • Molecular Sequence Data
  • Mutagenesis
  • Point Mutation
  • Protein Conformation
  • Protein Interaction Mapping
  • Protein Structure, Tertiary
  • Proto-Oncogene Proteins / metabolism*
  • Sequence Homology, Amino Acid
  • Signal Transduction
  • rap1 GTP-Binding Proteins / metabolism*

Substances

  • Integrins
  • KRIT1 Protein
  • KRIT1 protein, human
  • Microtubule-Associated Proteins
  • Proto-Oncogene Proteins
  • GTP Phosphohydrolases
  • rap1 GTP-Binding Proteins

Supplementary concepts

  • Familial cerebral cavernous malformation

Associated data

  • PDB/4DXA