Interactions between aldehyde derivatives and the aldehyde binding site of bacterial luciferase

✍ Scribed by Jockers, Ralf ;Ziegler, Torsten ;Schmid, Rolf D.

• Publisher: John Wiley and Sons
• Year: 1995
• Weight: 542 KB
• Volume: 10
• Category: Article
• ISSN: 0884-3996
• DOI: 10.1002/bio.1170100104

✦ Synopsis

The interaction o f triazine aldehydes w i t h the aldehyde binding site o f bacterial luciferases was investigated using a series of triazine aldehydes with different aldehyde chain length, and substituents on the s-triazine ring. Substrate activity was determined using luciferase from Photobacterium fischeri and Vibrio harveyi in a dithionite-based luciferases assay. The chain length optimum was determined for t w o triazine aldehyde classes t o be C-10 and C-11, respectively. Only the substrate activity o f 10-(4-chloro-6-methylthio-s-triazine-2-yl)aminodecanal (5) was as high as ndecanal, the reference aldehyde. A l l other triazine derivatives reduced light emission, probably by hindered binding o f the substrates. The degree o f activity reduction correlated with the volume o f the triazine ring moiety. The triazine moiety volume o f compound 5 was estimated t o be 200 x m3. Triazine aldehydes which showed reduced light emission had an estimated volume o f 228 x m3 or greater. All triazine aldehydes showed approximately 10-fold lower activities for Vibrio harveyi than for Photobacterium fischeri luciferase. Substrate specificity was the same for both luciferases. A schematic superposition o f quinone aldehydes and triazine aldehydes which showed substrate activities equivalent t o n-decanal, indicated potential interaction sites o f aldehyde substrates with the aldehyde binding site o f bacterial luciferases. The in vivo relevance o f the results is discussed.