Abstract
Photochemical crosslinking is a powerful method for studying all types of protein-nucleic acids interactions. In particular UV-induced crosslinking has been successfully applied to the study of protein-DNA interactions (e.g., ref. , see Chapters 23– 26 and 43). Ultraviolet (UV) light is a zero-length crosslinking agent. It is therefore not subject to the steric problems that can be associated with chemical crosslinking agents and provides strong evidence for close protein-DNA interactions. However, crosslinking with conventional UV-light sources requires exposure times ranging from minutes to several hours (e.g., see refs. , , ), permitting protein redistribution and the crosslinking of UV-damaged molecules. Because UV-laser irradiation is intense enough to induce crosslinking after very short exposure times, artifactual crosslinking can be avoided. The typically nanosecond or picosecond exposures times also allow UV-laser-induced crosslinking to be applied to study the intermediate states in rapid protein-DNA binding reactions (,).
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