Purification, N-terminal sequencing, partial characterization, crystallization and preliminary crystallographic analysis of two glycosylated serine proteinases from Agkistrodon acutus venom.

Zhongliang Zhu; Ping Gong; Maikun Teng; Liwen Niu

doi:10.1107/s0907444902023375

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PubMed · 12595722

Purification, N-terminal sequencing, partial characterization, crystallization and preliminary crystallographic analysis of two glycosylated serine proteinases from Agkistrodon acutus venom.

Zhongliang Zhu·Ping Gong·Maikun Teng·Liwen Niu

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Abstract

AaV-SP-I and AaV-SP-II, two glycosylated serine proteinases from Agkistrodon acutus venom with fibrinogenolysis and esterolysis activities, have been purified to homogeneity by three-step ion-exchange chromatography. Estimated by SDS-PAGE, the molecular weights of AaV-SP-I and AaV-SP-II are about 32 and 31 kDa under reducing conditions and 26 and 25 kDa under non-reducing conditions, respectively. The first 24 N-terminal amino-acid residues are the same in both sequences and display a high homology with those of several snake-venom serine proteinases. However, the proteins possess obviously distinct carbohydrate contents. Using the conventional hanging-drop vapour-diffusion method, single crystals of both enzymes were grown that were suitable for X-ray diffraction analysis. The crystals of AaV-SP-I and AaV-SP-II belong to space groups P2(1)2(1)2(1) and C2, respectively. In each case there is only one molecule in the asymmetric unit.

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Agkistrodon Amino Acid Sequence Ancrod Animals Carbohydrates Chromatography, Ion Exchange Crystallization Crystallography, X-Ray Electrophoresis, Polyacrylamide Gel Molecular Sequence Data Molecular Weight X-Ray Diffraction Journal Article Research Support, Non-U.S. Gov't

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Zhongliang Zhu, Ping Gong, Maikun Teng, Liwen Niu. 2003-02-21. Purification, N-terminal sequencing, partial characterization, crystallization and preliminary crystallographic analysis of two glycosylated serine proteinases from Agkistrodon acutus venom.. https://doi.org/10.1107/s0907444902023375

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