Activation of a bacterial lipase by its chaperone.
The gene lipA of Pseudomonas cepacia DSM 3959 encodes a prelipase from which a signal peptide is cleaved during secretion, producing a mature extracellular lipase. Expression of lipase in several heterologous hosts depends on the presence of another gene, limA, in cis or in trans. Lipase protein has been overproduced in Escherichia coli in the presence and absence of the lipase modulator gene limA. Therefore, limA is not required for the transcription of lipA or for the translation of the lipA mRNA. However, no lipase activity is observed in the absence of limA. limA has been overexpressed and encodes a 33-kDa protein, Lim. If lipase protein is denatured in 8 M urea and the urea is removed by dialysis, lipase activity is quantitatively recovered provided Lim protein is present during renaturation. Lip and Lim proteins form a complex precipitable either by an anti-lipase or anti-Lim antibody. The Lim protein has therefore the properties of a chaperone.
Hobson
A H
AH
Department of Genetics, Trinity College, Dublin, Ireland.
Buckley
C M
CM
Aamand
J L
JL
Jørgensen
S T
ST
Diderichsen
B
B
McConnell
D J
DJ
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D016956:Burkholderia cepacia / Q000201:enzymology* / Q000235:genetics; D003001:Cloning, Molecular; D004789:Enzyme Activation; D004792:Enzyme Precursors / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005798:Genes, Bacterial; D005809:Genes, Regulator; D007700:Kinetics; D008049:Lipase / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids; D014176:Protein Biosynthesis; D021382:Protein Sorting Signals / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D004792:Enzyme Precursors; D021382:Protein Sorting Signals; D012333:RNA, Messenger; D011994:Recombinant Proteins; D008049:Lipase