Biochemical and antigenic characterization of a new dipeptidyl-peptidase isolated from Aspergillus fumigatus
Biochemical and antigenic characterization of a new dipeptidyl-peptidase isolated from Aspergillus fumigatus
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DOI：
10.1074/jbc.272.10.6238
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发表时间：
1997-03-07
影响因子：
4.8
通讯作者：
Latge, JP
中科院分区：
文献类型：
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作者：
Beauvais, A;Monod, M;Latge, JP
从烟曲霉(Aspergillusfumgatus)培养基中分离纯化出一种新的二肽基肽酶(DPP V)。这是首次报道一种分泌型二肽基肽酶。该酶的相对分子质量为88 kDa，含有约9 kDa的N-连接碳水化合物，二肽基肽酶的表达和分泌随生长条件的不同而不同，当培养基中只含有蛋白质或蛋白质水解物而不含糖时，胞内和胞外表达水平最高。克隆了DPP V基因，并与其他真核生物的二肽基肽酶基因进行了序列同源性分析。与其他都在细胞内的二肽基肽酶不同，DPP V含有信号肽。与其他二肽基肽酶基因一样，DPP V显示了非经典丝氨酸蛋白酶催化位点的共同序列，在毕赤酵母中获得的天然和重组DPP V的生化性质是独特的，其特征是底物专一性，仅限于在中性pH条件下对X-丙氨酸、Ris-Ser和Ser-Tyr双肽的水解。此外，我们还证明了DPP V与用于诊断曲霉病的两种主要抗原之一相同。
A novel dipeptidyl-peptidase (DPP V) was purified from the culture medium of Aspergillus fumgatus. This is the first report of a secreted dipeptidyl-peptidase. The enzyme had a molecular mass of 88 kDa and contained approximately 9 kDa of N-linked carbohydrate, The expression and secretion of dipeptidyl-peptidase varied with the growth conditions; maximal intra- and extracellular levels were detected when the culture medium contained only proteins or protein hydrolysates in the absence of sugars. The gene of DPP V was cloned and showed significant sequence homology to other eukaryotic dipeptidyl-peptidase genes. Unlike the other dipeptidyl-peptidases, which are all intracellular, DPP V contained a signal peptide. Like the genes of other dipeptidyl-peptidases, that of DPP V displayed the consensus sequences of the catalytic site of the nonclassical serine proteases, The biochemical properties of native and recombinant DPP V obtained in Pichia pastoris were unique and were characterized by a substrate specificity limited to the hydrolysis of X-Ala, Ris-Ser, and Ser-Tyr dipeptides at a neutral pH optimum. In addition, we showed that DPP V is identical to one of the two major antigens used for the diagnosis of aspergillosis.