PMID: 8606184Apr 1, 1996Paper

Identification of the catalytic triad of the lipase/acyltransferase from Aeromonas hydrophila

Journal of Bacteriology
M J Brumlik, J T Buckley

Abstract

Aeromonas hydrophila secretes a lipolytic enzyme that has several properties in common with the mammalian enzyme lecithin-cholesterol acyltransferase. We have recently shown that it is a member of a newly described group of proteins that contain five similar blocks of sequence arranged in the same order in their primary structures (C. Upton and J. T. Buckley, Trends Biochem. Sci. 233:178-179, 1995). Assuming that, like other lipases, these enzymes have a Ser-Asp-His catalytic triad, we used these blocks to predict which aspartic acid and histidine would be at the active site of the Aeromonas enzyme. Targeted residues were replaced with other amino acids by site-directed mutagenesis, and the effects on secretion and activity were assessed. Changing His-291 to asparagine completely abolished enzyme activity, although secretion by the bacteria was not affected. Only very small amounts of the D116N mutant appeared in the culture supernatant, likely because it is sensitive to periplasmic proteases it encounters en route. Assays of crude preparations containing this variant showed no detectable enzyme activity. We conclude that, together with Ser-16, which we have identified previously, Asp-116 and His-291 compose the catalytic triad...Continue Reading

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Citations

Jun 22, 2000·Proceedings of the National Academy of Sciences of the United States of America·E A Miao, S I Miller
Feb 11, 2003·Molecular Plant-microbe Interactions : MPMI·Pablo VinuesaDietrich Werner
Dec 17, 2008·Future Microbiology·Sigrid D AuweterB Brett Finlay
Jan 9, 2010·Bioscience, Biotechnology, and Biochemistry·Atsushi MasayamaRyuichi Moriyama
Jun 25, 2010·Journal of Computational Chemistry·Xian-Hui WuYun-Dong Wu
May 16, 2008·Journal of Lipid Research·Akash DasLawrence L Rudel
Nov 21, 2007·Nature Reviews. Microbiology·Andrea HaragaSamuel I Miller
Jul 18, 2017·The Plant Journal : for Cell and Molecular Biology·Tam N T TranTimothy P Durrett
Nov 28, 2017·Scientific Reports·Christina LangAntje Flieger
May 18, 2017·Molecular Microbiology·Nikolai S ProkhorovAndrey V Letarov
Apr 6, 2019·Acta Crystallographica. Section F, Structural Biology Communications·Florian BleffertFilip Kovacic
Dec 31, 2019·International Journal of Analytical Chemistry·Tomas ValekMiroslav Pohanka
Dec 30, 2011·Applied Biochemistry and Biotechnology·Ramachandran Sivaramakrishnan, Karuppan Muthukumar
May 19, 2006·Extremophiles : Life Under Extreme Conditions·Maryam Goudarzi, Timothy L Born
Sep 24, 2005·Infection and Immunity·Maikke B OhlsonSamuel I Miller
Nov 13, 1999·Journal of Bacteriology·S WilhelmK E Jaeger
Aug 24, 2012·Plant Physiology·Panneerselvam VijayarajRam Rajasekharan
Jun 10, 1998·Microbiology and Molecular Biology Reviews : MMBR·P StanleyC Hughes
Jul 31, 2016·Applied and Environmental Microbiology·Annika RöttigAlexander Steinbüchel
Dec 8, 2004·Archives of Biochemistry and Biophysics·Corey M JohnsonPaul F Cook
Nov 4, 2004·Progress in Lipid Research·Casimir C AkohJei-Fu Shaw
Oct 7, 2008·Bioresource Technology·N C MhetrasD V Gokhale

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