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

Abstract

CitationPatel TR, Nikodemus D, Besong TMD, Reuten R, Meier M, et al. (2015) Biophysical analysis of a lethal laminin alpha-1 mutation reveals altered self-interaction. Matrix Biology. AbstractLaminins 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.

The occhiolino (occ) mutant Zebrafish, a model for development of the optical function in the biological lens

“…In the developing lens, the capsule is a basement membrane for the attachment of the basal surface of epithelial cells during the synchronized proliferation, migration, and elongation to form layers of transparent lens fibers organized precisely into the optical element of the Zebrafish visual system. As a key component of the basement membrane, laminin is a marker for the lens capsule (Parmigiani and McAvoy, ; Menko et al, ; Norose et al, ; Yan et al, ; Lee and Gross, ; Danysh and Duncan, ; Kwan, ; James et al, ; Patel et al, ). Laminin mutants disrupt basement membrane structure, resulting in abnormal lens development in Zebrafish as early as 3 dpf, with some mutations causing complete loss of the lens as early at 7 dpf (Parmigiani and McAvoy, ; Semina et al, ; Zinkevich et al, ; Pathania et al, ; Patel et al, ).…”

“…As a key component of the basement membrane, laminin is a marker for the lens capsule (Parmigiani and McAvoy, ; Menko et al, ; Norose et al, ; Yan et al, ; Lee and Gross, ; Danysh and Duncan, ; Kwan, ; James et al, ; Patel et al, ). Laminin mutants disrupt basement membrane structure, resulting in abnormal lens development in Zebrafish as early as 3 dpf, with some mutations causing complete loss of the lens as early at 7 dpf (Parmigiani and McAvoy, ; Semina et al, ; Zinkevich et al, ; Pathania et al, ; Patel et al, ). The involvement of laminin in the formation of the basement membrane was the basis for using laminin antibodies to label the lens capsule in wt and occ eyes (Pathania et al, ; Pathania et al, ).…”

“…Movies 1-3.) in abnormal lens development in Zebrafish as early as 3 dpf, with some mutations causing complete loss of the lens as early at 7 dpf (Parmigiani and McAvoy, 1991;Semina et al, 2006;Zinkevich et al, 2006;Pathania et al, 2014;Patel et al, 2016). The involvement of laminin in the formation of the basement membrane was the basis for using laminin antibodies to label the lens capsule in wt and occ eyes (Pathania et al, 2014;Pathania et al, 2016).…”

A Hilly path through the thermodynamics and statistical mechanics of protein solutions

“…The methodologies and interpretations of experimental data that were developed by him and his many collaborators continue to be applied to a wide range of problems of biological and medical significance, especially in the field of protein-protein interactions. A multitude of structure-function studies, recently ranging from the molecular basis of murine olefaction (Portman et al 2014), through the heterodimeric character of a plant immune receptor (Williams et al 2014), to a lethal mutation in the laminin alpha-1 gene (Patel et al 2015), has demonstrated the importance of the thermodynamic approach to significant molecular biological problems. It can only be anticipated that the influence of Don's work and its application will still be felt many decades into the future.…”

Nanoscale Assembly of High-Mobility Group AT-Hook 2 Protein with DNA Replication Fork

“…SEC-purified samples were subjected to 0.1 mm filtration before equilibrating at 20 C for 5 min for DLS analysis. The hydrodynamic radius (R h ) distributions were obtained using a Nano-S Zetasizer (Malvern, Toronto, ON, Canada) at concentrations between 1 and 10 mg/mL as described previously (29)(30)(31). SV experiments were performed as previously described (30,32)…”