Hobson A H, Buckley C M, Jørgensen S T, Diderichsen B, McConnell D J
Department of Genetics, Trinity College, Dublin 2, Ireland.
J Biochem. 1995 Sep;118(3):575-81. doi: 10.1093/oxfordjournals.jbchem.a124948.
The lipA gene of Pseudomonas cepacia DSM3959 requires a downstream gene, limA, in oder to express lipase activity. The product of the lim gene, LimA, is a molecular chaperone required during the folding of lipase in oder for the lipase to adopt an active conformation. The lipase and LimA proteins have been shown to form a complex precipitable with either an anti-lipase or anti-LimA antibody. LimA has been shown to form a 1:1 complex with with prelipase and lipase isolated from "natural" P. cepacia system. The mature lipase (lacking its signal peptide) has been expressed in the presence and absence of LimA in Escherichia coli. LimA can activate mature lipase during a urea denaturation-renaturation experiment, indicating that the signal peptide is not required for the lipase to be activated by LimA. The effects of various reagents on the renaturation of lipase from 8 M urea have been examined. We propose a mechanism for the function of the LimA chaperone during the production of active extracellular lipase.
洋葱伯克霍尔德菌DSM3959的lipA基因需要下游基因limA才能表达脂肪酶活性。lim基因的产物LimA是脂肪酶折叠过程中所需的分子伴侣,以使脂肪酶采用活性构象。脂肪酶和LimA蛋白已被证明能形成一种可被抗脂肪酶或抗LimA抗体沉淀的复合物。LimA已被证明能与从“天然”洋葱伯克霍尔德菌系统中分离出的前脂肪酶和脂肪酶形成1:1复合物。成熟脂肪酶(缺少其信号肽)已在大肠杆菌中分别在有和没有LimA的情况下表达。在尿素变性-复性实验中,LimA可以激活成熟脂肪酶,这表明脂肪酶被LimA激活不需要信号肽。已经研究了各种试剂对8M尿素中脂肪酶复性的影响。我们提出了一种在活性细胞外脂肪酶产生过程中LimA分子伴侣功能的机制。