CRYSTAL-STRUCTURES OF NATIVE AND INHIBITED FORMS OF HUMAN CATHEPSIN-D - IMPLICATIONS FOR LYSOSOMAL TARGETING AND DRUG DESIGN
CRYSTAL-STRUCTURES OF NATIVE AND INHIBITED FORMS OF HUMAN CATHEPSIN-D - IMPLICATIONS FOR LYSOSOMAL TARGETING AND DRUG DESIGN
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DOI：
10.1073/pnas.90.14.6796
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发表时间：
1993-07-15
影响因子：
11.1
通讯作者：
ERICKSON, JW
中科院分区：
文献类型：
--
作者：
BALDWIN, ET;BHAT, TN;ERICKSON, JW
组织蛋白酶D（EC 3.4.23.5）是一种溶酶体蛋白酶，被怀疑在蛋白质催化、抗原加工、退行性疾病和乳腺癌进展中起重要作用。在2.5埃分辨率下测定组织蛋白酶D和胃蛋白酶抑制剂复合物的晶体结构，为这种双链N-糖基化天冬氨酸蛋白酶的抑制剂结合和溶酶体靶向提供了见解。胃蛋白酶抑制剂结合到根霉胃蛋白酶和与人的肾素抑制剂复合物的复合物的结构的比较，揭示了亚位点结构和通道-酶相互作用的差异，这与亲和力差异和结构-活性关系是一致的，并建议微调组织蛋白酶D抑制剂的特异性的策略。诱变研究已经确定了一个磷酸转移酶识别区域，该区域是寡糖磷酸化所需的，但距离Asn-70的N-结构域糖基化位点32埃。组织蛋白酶D晶体结构的电子密度表明存在从Asn-70向Lys-203延伸的N-连接的寡糖，其是磷酸转移酶识别区域的关键组分，从而为磷酸转移酶如何识别蛋白质表面上明显遥远的位点提供了结构解释。
Cathepsin D (EC 3.4.23.5) is a lysosomal protease suspected to play important roles in protein catabolism, antigen processing, degenerative diseases, and breast cancer progression. Determination of the crystal structures of cathepsin D and a complex with pepstatin at 2.5 angstrom resolution provides insights into inhibitor binding and lysosomal targeting for this two-chain, N-glycosylated aspartic protease. Comparison with the structures of a complex of pepstatin bound to rhizopuspepsin and with a human renin-inhibitor complex revealed differences in subsite structures and inhibitor-enzyme interactions that are consistent with affinity differences and structure-activity relationships and suggest strategies for fine-tuning the specificity of cathepsin D inhibitors. Mutagenesis studies have identified a phosphotransferase recognition region that is required for oligosaccharide phosphorylation but is 32 angstrom distant from the N-domain glycosylation site at Asn-70. Electron density for the crystal structure of cathepsin D indicated the presence of an N-linked oligosaccharide that extends from Asn-70 toward Lys-203, which is a key component of the phosphotransferase recognition region, and thus provides a structural explanation for how the phosphotransferase can recognize apparently distant sites on the protein surface.