Abstract
A Drosophila laminin that has the chain composition alpha5 beta1 gamma1, relative to mammalian laminins, bound human and mouse nidogen almost as strongly as mouse laminin-1 (alpha1 beta1 gamma1) in solid-phase assays, and had only a fourfold lower affinity in a radioligand competition test. This is due to a short, highly conserved sequence that occurs in both laminin gamma1 chains and which binds nidogen. When the single conservative amino acid difference between the two sequences (Tyr-->His) was introduced into the mouse laminin binding module gamma1 III4 it failed to cause any change of binding. A high affinity between Drosophila laminin and mouse nidogen resulted in the formation of a stable complex in solution. Drosophila laminin also bound to the mouse heparan sulfate proteoglycan perlecan and the formation of this complex was inhibited by heparin, but not by chondroitin sulfate. In addition, a weaker connection between the core protein of mouse perlecan and Drosophila laminin can be mediated through nidogen. Elastase and other proteases degraded Drosophila laminin to a restricted number of larger fragments (40-300 kDa), almost all of which were bound to a heparin affinity column. Three fragments could be displaced at low salt concentration and were derived from the short arms of the Drosophila laminin, as shown by sequence analysis. A more strongly bound 50-kDa fragment apparently comprised the globular domains LG2 and LG3 derived from the C-terminal part of its alpha chain. Therefore, Drosophila laminin and mouse laminin-1 differ in certain aspects of protease stability and heparin-binding sites that, in part, can be attributed to their different alpha chains. The data suggest the existence of a nidogen analog and heparan sulfate proteoglycans in Drosophila, which remain to be identified.
