Purification and crystallization of precursors and autoprocessed enzymes of Flavobacterium glycosylasparaginase:: an N-terminal nucleophile hydrolase
Purification and crystallization of precursors and autoprocessed enzymes of Flavobacterium glycosylasparaginase:: an N-terminal nucleophile hydrolase
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DOI：
10.1107/s0907444999011798
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发表时间：
1999-11-01
期刊：
影响因子：
--
通讯作者：
Guo, HC
中科院分区：
文献类型：
--
作者：
Cui, T;Liao, PH;Guo, HC
糖基天冬酰胺酶(GA)是一组新的蛋白质，通过自身催化的多肽键从单链前体裂解而产生水解酶活性所需的两个亚基。野生型GA前体会自发地分解成α和β亚基。本文报道了以单链前体和成熟(自动加工)形式纯化脑膜败血黄杆菌赤霉素A的策略。重组蛋白以不同的空间群结晶：前体酶为P1，成熟酶为P2(1)。在实验室X射线照射下，前驱体晶体的衍射率达到1.9埃。
Glycosylasparaginase (GA) represents a novel group of proteins that are activated by self-catalyzed peptide-bond cleavage from a single-chain precursor to yield the two subunits required for hydrolase activity. The wild-type GA precursor autoproteolyzes spontaneously into alpha and beta subunits. Strategies are reported here for purification to homogeneity of GA from Flavobacterium meningosepticum in both single-chain precursor and mature (autoprocessed) forms. The recombinant proteins crystallize in different space groups: P1 and P2(1) for the precursor and mature enzymes, respectively. The precursor crystals diffract to 1.9 Angstrom resolution with laboratory X-ray radiation.