Fulcher I S, Pappin D J, Kenny A J
Biochem J. 1986 Nov 15;240(1):305-8. doi: 10.1042/bj2400305.
Endopeptidase-24.11 (EC 3.4.24.11), a widely distributed ectoenzyme, was isolated from pig kidneys by detergent solubilization of membranes and immuno-affinity chromatography. In all, 12 preparations of the enzyme were submitted to solid-phase sequencing, yielding a consensus sequence of 25 amino acid residues of the N-terminal segment. Some samples were treated with either trypsin or Staphylococcus aureus V8 proteinase before sequencing. There were four lysine and one arginine residues in the first nine positions. This segment was susceptible to hydrolysis by trypsin and, in some samples, to endogenous proteinases. From residue 19 onwards, the sequence became intensely hydrophobic. There was a striking homology with the N-terminal sequence of pro-sucrase-isomaltase. From Lys7 to Leu20 there were seven identical amino acid residues and four conservative substitutions. We suggest that endopeptidase-24.11 is topologically similar to this glycosidase, the N-terminus at the cytoplasmic face and hydrophobic segment serving the roles of both signal peptide and hydrophobic anchor.
内肽酶-24.11(EC 3.4.24.11)是一种广泛分布的胞外酶,通过用去污剂溶解膜并进行免疫亲和层析从猪肾中分离得到。总共对12份该酶制剂进行了固相测序,得到了N端片段25个氨基酸残基的共有序列。一些样品在测序前用胰蛋白酶或金黄色葡萄球菌V8蛋白酶处理。在前九个位置有四个赖氨酸残基和一个精氨酸残基。该片段易被胰蛋白酶水解,在一些样品中也易被内源性蛋白酶水解。从第19位残基开始,序列变得高度疏水。它与蔗糖酶-异麦芽糖酶原的N端序列有显著同源性。从Lys7到Leu20有七个相同的氨基酸残基和四个保守替换。我们认为内肽酶-24.11在拓扑结构上与这种糖苷酶相似,其N端位于细胞质面,疏水片段兼具信号肽和疏水锚的作用。