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首页> 外文期刊>Journal of Molecular Biology >Crystal structure of N-acyl-D-glucosamine 2-epimerase from porcine kidney at 2.0 angstrom resolution
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Crystal structure of N-acyl-D-glucosamine 2-epimerase from porcine kidney at 2.0 angstrom resolution

机译：猪肾脏N-酰基-D-氨基葡萄糖2-表异构酶的晶体结构，分辨率为2.0埃

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摘要

The X-ray crystallographic structure of N-acyl-D-glucosamine 2-epimerase (AGE) from porcine kidney, which has been identified to be a renin-binding protein (RnBP), was determined by the multiple isomorphous replacement method and refined at 2.0 Angstrom resolution with a final R-factor of 16.9% for 15 to 2.0 Angstrom resolution data. The refined structure of AGE comprised 804 amino acid residues tone dimer) and 145 water molecules. The dimer of AGE had an asymmetric unit with approximate dimensions 46 Angstrom x 48 Angstrom x 96 Angstrom. The AGE monomer is composed of an alpha (6)/alpha (6)-barrel, the structure of which is found in glucoamylase and cellulase. One side of the AGE alpha (6)/alpha (6)-barrel structure comprises long loops containing five short beta -sheets, and contributes to the formation of a deep cleft shaped like a funnel. The putative active-site pocket and a possible binding site for the substrate N-acetyl-D-glucosamine (GlcNAc) were found in the cleft. The other side of the alpha (6)/alpha (6)-barrel comprises short loops and contributes to the dimer formation. At the dimer interface, which is composed of the short loops and alpha -helices of the subunits, five strong ion-pair interactions were observed, which play a major role in the dimer assembly. This completely ruled out the previously accepted hypothesis that the formation of the RnBP homodimer and RnBP-renin heterodimer requires the leucine zipper motif present in RnBP. (C) 2000 Academic Press. [References: 35]
机译：猪肾脏的N-酰基-D-氨基葡萄糖2-表异构酶（AGE）的X射线晶体结构通过多重同构置换法确定，并在60℃下精制，该结构被鉴定为肾素结合蛋白（RnBP）。对于15至2.0埃分辨率的数据，分辨率为2.0埃，最终R因子为16.9％。 AGE的精制结构包含804个氨基酸残基（色调二聚体）和145个水分子。 AGE的二聚体具有不对称单元，其近似尺寸为46埃×48埃×96埃。 AGE单体由α（6）/α（6）桶组成，其结构存在于葡糖淀粉酶和纤维素酶中。 AGEα（6）/α（6）-桶结构的一侧包括长环，该环包含五个短的β-折叠，并有助于形成漏斗状的深裂。在裂缝中发现了假定的活性位点口袋和底物N-乙酰基-D-葡萄糖胺（GlcNAc）的可能结合位点。 α（6）/α（6）-桶的另一侧包括短环，并有助于形成二聚体。在由短环和亚基的α-螺旋组成的二聚体界面上，观察到五个强离子对相互作用，这在二聚体组装中起主要作用。这完全排除了先前公认的假设，即RnBP同二聚体和RnBP-肾素异二聚体的形成需要RnBP中存在亮氨酸拉链基序。 （C）2000学术出版社。 [参考：35]

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