Chemical crosslinking of cell membranes.
The complexity of cell membranes makes the resolution of their macromolecular topology one of the more challenging problems in modern molecular and cellular biochemistry. Despite the difficulties inherent in any such analysis, a surprisingly simple yet powerful approach exists that has consistently yielded valuable results. This method is chemical crosslinking, in which cell membranes are treated with crosslinking reagents (usually bifunctional) which produce covalent linkages between membrane components. The resultant complexes are usually then separated and identified by electrophoresis. This review is intended to provide a guide to the investigator who is unfamiliar with this approach. The overall strategy of crosslinking is discussed including selection of reagents, conditions to optimize crosslinking and the cleavage of crosslinked complexes to regenerate the original target for identification purposes. The crosslinking of biological membranes is then reviewed with special emphasis on recent advances including macromolecular photoaffinity labeling, kinetic analysis to probe symmetry properties and potential artifacts that may complicate interpretation of results. Examples of specific applications of crosslinking to membranes are presented in tabular form. The final portion of the review discusses the synthesis and properties of the most widely employed crosslinking reagents. Available reagents are summarized in a series of comprehensive tables. It is hoped that our discussion will provide the uninitiated investigator with sufficient information to ascertain the applicability of chemical crosslinking to particular areas of interest.
D000345:Affinity Labels; D001386:Azides; D002462:Cell Membrane / Q000032:analysis*; D055598:Chemical Phenomena; D002621:Chemistry; D003432:Cross-Linking Reagents; D007096:Imidoesters; D007700:Kinetics; D010777:Photochemistry; D013388:Succinimides; D013439:Sulfhydryl Reagents
D000345:Affinity Labels; D001386:Azides; D003432:Cross-Linking Reagents; D007096:Imidoesters; D013388:Succinimides; D013439:Sulfhydryl Reagents