Quantification of coral heat shock proteins from individual coral polyps

Author Posting. © Inter-Research, 2009. This article is posted here by permission of Inter-Research for personal use, not for redistribution. The definitive version was published in marine Ecology Progress Series 376 (2009): 123-132, doi:10.3354/meps07812.The induction and regulation of heat shock proteins (hsps) is a significant defense mechanism that can preserve metabolic function and foster recovery from short-term stress events. Present coral sampling methodologies that involve hsp analysis often require the harvesting of large samples of live coral colonies that may already be stressed or in poor health. In the present study, 3 novel protocols were developed to: (1) extract single coral polyps, minimizing colony trauma; (2) purify protein from single coral polyps (approximately 12 mm3); and (3) develop a more sensitive protein quantification method. The preliminary testing of 5 separate protein preparation methods resulted in a range of total protein yields from 47 to 77 µg coral polyp–1. The optimized methods were able to recover, on average, 44 ± 12 µg protein polyp–1 (n = 20). Subsequent SDS-PAGE and immunoblotting analysis of single coral polyps resolved as little as 87 pg hsp70 coral polyp–1. This minimally invasive sampling protocol reduces coral damage and, thus, reduces stress and diseases caused by sampling.This research was funded by a grant from the National Oceanic and Atmospheric Administration’s Undersea Research Center at the University of North Carolina at Wilmington pursuant to NOAA Award Number NA03OAR4300088