Bode W, Wei A Z, Huber R, Meyer E, Travis J, Neumann S
EMBO J. 1986 Oct;5(10):2453-8. doi: 10.1002/j.1460-2075.1986.tb04521.x.
Orthorhombic crystals diffracting beyond 1.7 A resolution, have been grown from the stoichiometric complex formed between human leukocyte elastase (HLE) and the third domain of turkey ovomucoid inhibitor (OMTKY3). The crystal and molecular structure has been determined with the multiple isomorphous replacement technique. The complex has been modeled using the known structure of OMTKY3 and partial sequence information for HLE, and has been refined. The current crystallographic R-value is 0.21 for reflections from 25 to 1.8 A resolution. HLE shows the characteristic polypeptide fold of trypsin-like serine proteinases and consists of 218 amino acid residues. However, several loop segments, mainly arranged around the substrate binding site, have unique conformations. The largest deviations from the other vertebrate proteinases of known spatial structure are around Cys168. The specificity pocket is constricted by Val190, Val216 and Asp226 to preferentially accommodate medium sized hydrophobic amino acids at P1. Seven residues of the OMTKY3-binding segment are in specific contact with HLE. This interaction and geometry around the reactive site are similar as observed in other complexes. It is the first serine proteinase glycoprotein analysed, having two sugar chains attached to Asn159 and to residue 109.
从人白细胞弹性蛋白酶(HLE)与火鸡卵类粘蛋白抑制剂第三结构域(OMTKY3)形成的化学计量复合物中生长出了能在1.7埃分辨率以上衍射的正交晶体。晶体和分子结构已通过多重同晶置换技术确定。利用已知的OMTKY3结构和HLE的部分序列信息对该复合物进行了建模,并进行了优化。对于25至1.8埃分辨率的反射,当前晶体学R值为0.21。HLE显示出胰蛋白酶样丝氨酸蛋白酶的特征性多肽折叠,由218个氨基酸残基组成。然而,几个环段,主要围绕底物结合位点排列,具有独特的构象。与已知空间结构的其他脊椎动物蛋白酶最大的偏差出现在Cys168周围。特异性口袋被Val190、Val216和Asp226限制,优先容纳P1位点的中等大小疏水氨基酸。OMTKY3结合段的七个残基与HLE有特异性接触。这种相互作用以及活性位点周围的几何结构与在其他复合物中观察到的相似。这是第一个被分析的丝氨酸蛋白酶糖蛋白,有两条糖链分别连接在Asn159和第109位残基上。