重组大鼠肌醇-1,4,5-三磷酸3-激酶B的表达表明其N端具有调节作用。
Expression of recombinant rat myo-inositol 1,4,5-trisphosphate 3-kinase B suggests a regulatory role for its N-terminus.
作者信息
Thomas S, Soriano S, d'Santos C, Banting G
机构信息
Department of Biochemistry, University of Bristol, U.K.
出版信息
Biochem J. 1996 Nov 1;319 ( Pt 3)(Pt 3):713-6. doi: 10.1042/bj3190713.
Abstract
We have expressed rat myo-inositol 1,4,5-trisphosphate (IP3) 3-kinase B as both a full-length, recombinant, non-fusion protein and a full-length, recombinant, fusion protein with maltose-binding protein (MBP) in Escherichia coli. The fusion protein with MBP is soluble, binds calmodulin and is enzymically active whereas the non-fusion protein is insoluble and does not bind calmodulin unless co-expressed with bacterial chaperone proteins (either GroES and GroEL, or DnaK, DnaJ and GrpE). However, soluble, calmodulin-binding non-fusion IP3 3-kinase B is enzymically inactive. The catalytic domain of the enzyme has previously been shown to reside near the C-terminus; the results we present suggest an auto-regulatory role for the N-terminus.
摘要
我们已在大肠杆菌中表达了大鼠肌醇1,4,5-三磷酸（IP3）3-激酶B，其形式包括全长重组非融合蛋白以及与麦芽糖结合蛋白（MBP）的全长重组融合蛋白。与MBP的融合蛋白是可溶的，能结合钙调蛋白且具有酶活性，而该非融合蛋白是不溶的，除非与细菌伴侣蛋白（GroES和GroEL，或DnaK、DnaJ和GrpE）共表达，否则不结合钙调蛋白。然而，可溶的、结合钙调蛋白的非融合IP3 3-激酶B没有酶活性。此前已表明该酶的催化结构域位于C端附近；我们给出的结果表明N端具有自动调节作用。
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