Biophysical analysis of a lethal laminin alpha-1 mutation reveals altered self-interaction

Research output: Contribution to journalArticlepeer-review

Authors

  • Denise Nikodemus
  • Tabot M.d. Besong
  • Raphael Reuten
  • Markus Meier
  • Stephen E. Harding
  • Donald J. Winzor
  • Manuel Koch
  • Jörg Stetefeld

Colleges, School and Institutes

Abstract

Laminins are key basement membrane molecules that influence several biological activities and are linked to a number of diseases. They are secreted as heterotrimeric proteins consisting of one α, one β, and one γ chain, followed by their assembly into a polymer-like sheet at the basement membrane. Using sedimentation velocity, dynamic light scattering, and surface plasmon resonance experiments, we studied self-association of three laminin (LM) N-terminal fragments α-1 (hLM α-1 N), α-5 (hLM α-5 N) and β-3 (hLM β-3 N) originating from the short arms of the human laminin αβγ heterotrimer. Corresponding studies of the hLM α-1 N C49S mutant, equivalent to the larval lethal C56S mutant in zebrafish, have shown that this mutation causes enhanced self-association behavior, an observation that provides a plausible explanation for the inability of laminin bearing this mutation to fulfill functional roles in vivo, and hence for the deleterious pathological consequences of the mutation on lens function.

Details

Original languageEnglish
JournalMatrix Biology
Early online date26 Jul 2015
Publication statusE-pub ahead of print - 26 Jul 2015

Keywords

  • Analytical ultracentrifugation, CD spectroscopy, Dynamic light scattering, Extracellular matrix, Laminin short arms, Protein self-association, Surface plasmon resonance