Chaperone-mediated activation in vivo of a Pseudomonas cepacia lipase |
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Authors: | Jesper L Aamand Audrey H Hobson Catherine M Buckley Steen T Jørgensen Borge Diderichsen and David J McConnell |
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Institution: | (1) Department of Genetics, Trinity College, IE-Dublin 2, Ireland;(2) National Pharmaceutical Biotechnology Centre, Trinity College, IE-Dublin 2, Ireland;(3) Department of Bacterial Gene Technology, Novo Nordisk A/S, DK-2880 Bagsvaerd, Denmark |
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Abstract: | An extracellular Pseudomonas cepacia lipase, LipA, is inactive when expressed in the absence of the product of the limA gene. Evidence has been presented that LimA is a molecular chaperone. The lipA and limA genes have been cloned in separate and independently inducible expression systems in Escherichia coli. These systems were used to test the molecular chaperone hypothesis by investigating whether LimA could activate presynthesized prelipase and whether presynthesized LimA could activate newly synthesized prelipase. The results show that LimA cannot activate presynthesized prelipase and that presynthesized LimA can activate only a limited number of de novo synthesized prelipase molecules. Co-immunoprecipitation of prelipase/lipase with LimA generated a 1:1 complex of prelipase/lipase and LimA. The results suggest that a 1:1 complex of LipA and LimA is required for prelipase processing and secretion of active lipase. |
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Keywords: | Chaperone Pseudomonas cepacia lipase LimA |
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