Characterization of sodium dodecyl sulfate-resistant proteolytic activity in the hyperthermophilic archaebacterium Pyrococcus furiosus.

Applied and Environmental Microbiology
I I BlumentalsR M Kelly

Abstract

Cell extracts from Pyrococcus furiosus were found to contain five proteases, two of which (S66 and S102) are resistant to sodium dodecyl sulfate (SDS) denaturation. Cell extracts incubated at 98 degrees C in the presence of 1% SDS for 24 h exhibited substantial cellular proteolysis such that only four proteins could be visualized by amido black-Coomassie brilliant blue staining of SDS-polyacrylamide gels. The SDS-treated extract retained 19% of the initial proteolytic activity as represented by two proteases, S66 (66 kilodaltons [kDa]) and S102 (102 kDa). Immunoblot analysis with guinea pig sera containing antibodies against protease S66 indicated that S66 is related neither to S102 nor to the other proteases. The results of this analysis also suggest that S66 might be the hydrolysis product of a 200-kDa precursor which does not have proteolytic activity. The 24-h SDS-treated extract showed unusually thermostable proteolytic activity; the measured half-life at 98 degrees C was found to be 33 h. Proteases S66 and S102 were also resistant to denaturation by 8 M urea, 80 mM dithiothreitol, and 5% beta-mercaptoethanol. Purified protease S66 was inhibited by phenylmethylsulfonyl fluoride and diisopropyl fluorophosphate but not by ED...Continue Reading

References

Sep 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·H TowbinJ Gordon

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Citations

Jan 1, 1994·Antonie van Leeuwenhoek·R M Kelly, M W Adams
Dec 13, 2000·Proceedings of the National Academy of Sciences of the United States of America·X DuS H Kim
Nov 20, 2004·The Journal of Biological Chemistry·Abderrahim MalkiGilbert Richarme
Dec 11, 2003·The Journal of Biological Chemistry·James A OlzmannLih-Shen Chin
Jun 23, 2006·FEMS Microbiology Letters·Sylvain C P EschenlauerJeremy C Mottram
Apr 2, 2008·Annals of the New York Academy of Sciences·Tairo Oshima, Toshiyuki Moriya
Oct 16, 2007·Journal of Bacteriology·Jad AbdallahGilbert Richarme
Jan 1, 1995·World Journal of Microbiology & Biotechnology·C Leuschner, G Antranikian
Jul 1, 1995·Bio/technology·M W AdamsR M Kelly
May 22, 2002·Endocrinology·John J KopchickShigeru Okada
Sep 23, 2003·Protein Science : a Publication of the Protein Society·Yonghong ZhaoRobert O Fox
Apr 25, 1993·Biotechnology and Bioengineering·C B Pepper, H G Monbouquette
Oct 27, 2015·Applied and Environmental Microbiology·Dongheng MengBing Tang
Aug 1, 1994·Applied Biochemistry and Biotechnology·J G Morris
Nov 27, 2010·Journal of Industrial Microbiology & Biotechnology·Sreeja ChellappanM Chandrasekaran
Jun 1, 2015·Journal, Genetic Engineering & Biotechnology·Aysha KamranMustafa Kamal
Jun 13, 2016·Extremophiles : Life Under Extreme Conditions·Irina Shlaifer, Joanne L Turnbull
Jun 27, 2018·FEMS Microbiology Reviews·Christopher T StraubRobert M Kelly
Sep 7, 2017·Acta Crystallographica. Section D, Structural Biology·Steven B Larson, Alexander McPherson
Feb 17, 2017·Wiley Interdisciplinary Reviews. Systems Biology and Medicine·James A CountsRobert M Kelly
Jul 1, 1994·Bioorganic & Medicinal Chemistry·M W Adams, R M Kelly
Mar 1, 1993·Journal of Bacteriology·R N SchichoR M Kelly
Apr 1, 1992·Applied and Environmental Microbiology·L J SnowdenR M Kelly
Feb 5, 2003·Biochemical and Biophysical Research Communications·Abderrahim MalkiGilbert Richarme
Mar 1, 1997·Applied and Environmental Microbiology·M W BauerR M Kelly

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