Analysis of basement membrane self-assembly and cellular interactions with native and recombinant glycoproteins

Abstract

Publisher Summary This chapter discusses the biochemical and structural methods used to characterize binding interactions in both native and recombinant glycoproteins, and describe approaches that can be used to study cell-surface interactions and the relationships between matrix binding and cell-surface accumulation. Basement membrane formation is one of self-assembly in which both laminins and type IV collagens polymerize and bind to nidogen, perlecan, and, where presents, agrin. These extracellular matrices (ECMs) can be many microns thick, and only a small fraction of components may be close enough to the cell surface to be able to interact with a receptor. Basement membrane formation can be separated into several steps. These are (1) the synthesis of structural components and their folding to form active molecules; (2) secretion into the extracellular compartment; (3) adhesion of components to target cell surfaces; (4) self-assembly through polymerization and intercomponent bond formation to create an extracellular supramolecular complex; and (5) turnover, reflecting the balance of formation and degradation of assembled components that is an important part of tissue remodeling. A defect in any one of these steps may lead to the failure to develop or detect a basement membrane in a correct tissue location.