title: Biophysical analysis of a lethal laminin alpha-1 mutation reveals altered self-interaction.
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-1N), α-5 (hLM α-5N) and β-3 (hLM β-3N) originating from the short arms of the human laminin αβγ heterotrimer. Corresponding studies of the hLM α-1N 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.
authors.affiliation: Department of Chemistry, University of Manitoba, 144 Dysart Road, Winnipeg, Manitoba R3T 2N2, Canada; School of Biosciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, United Kingdom. Electronic address: t.r.patel@bham.ac.uk.
authors.forename: Trushar R
authors.initials: TR
authors.lastname: Patel
authors.affiliation: Institute for Dental Research and Oral Musculoskeletal Biology and Center for Biochemistry, Medical Faculty, University of Cologne, Cologne 50931, Germany.
authors.forename: Denise
authors.initials: D
authors.lastname: Nikodemus
authors.affiliation: National Center for Macromolecular Hydrodynamics, University of Nottingham, Sutton Bonington LE12 5RD, United Kingdom; Functional Nanomaterials Lab, King Abdullah University of Science and Technology (KAUST), Thuwal 23955-6900, Saudi Arabia.
authors.forename: Tabot M D
authors.initials: TMD
authors.lastname: Besong
authors.affiliation: Institute for Dental Research and Oral Musculoskeletal Biology and Center for Biochemistry, Medical Faculty, University of Cologne, Cologne 50931, Germany.
authors.forename: Raphael
authors.initials: R
authors.lastname: Reuten
authors.affiliation: Department of Chemistry, University of Manitoba, 144 Dysart Road, Winnipeg, Manitoba R3T 2N2, Canada.
authors.forename: Markus
authors.initials: M
authors.lastname: Meier
authors.affiliation: National Center for Macromolecular Hydrodynamics, University of Nottingham, Sutton Bonington LE12 5RD, United Kingdom.
authors.forename: Stephen E
authors.initials: SE
authors.lastname: Harding
authors.affiliation: School of Chemistry and Molecular Biosciences, University of Queensland, Brisbane, Queensland 4072, Australia.
authors.forename: Donald J
authors.initials: DJ
authors.lastname: Winzor
authors.affiliation: School of Biosciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, United Kingdom.
authors.forename: Manuel
authors.initials: M
authors.lastname: Koch
authors.affiliation: Department of Chemistry, University of Manitoba, 144 Dysart Road, Winnipeg, Manitoba R3T 2N2, Canada. Electronic address: jorg.stetefeld@ad.umanitoba.ca.
authors.forename: Jörg
authors.initials: J
authors.lastname: Stetefeld
mesh_terms: D001665:Binding Sites; D000067493:Dynamic Light Scattering; D005804:Genes, Lethal; D006801:Humans; D007797:Laminin / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D008958:Models, Molecular; D009154:Mutation; D011487:Protein Conformation; D055503:Protein Multimerization; D017434:Protein Structure, Tertiary; D020349:Surface Plasmon Resonance
chemical_list: D007797:Laminin; C084098:laminin A
keywords: Analytical ultracentrifugation; CD spectroscopy; Dynamic light scattering; Extracellular matrix; Laminin short arms; Protein self-association; Surface plasmon resonance