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High-Level Expression, Purification, Characterization and Structural Prediction of a Snake Venom Metalloproteinase Inhibitor in Pichia pastoris

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Abstract

Snake venom metalloproteinase inhibitor BJ46a is from the serum of the venomous snake Bothrops jararaca. It has been proven to possess the capacity to inhibit matrix metalloproteinases (MMPs), likely based on its structural similarity to MMPs. This report describes the successful expression, purification, and characterization of the recombinant protein BJ46a in Pichia pastoris. Purified recombinant protein BJ46a was found to inhibit MMPs. Structural modeling was completed and should provide the foundation for further functional research. To our knowledge, this is the first report on the large scale expression of BJ46a, and it provides promise as a method for generation of BJ46a and investigation of its potential use as a new drug for treatment of antitumor invasion and metastasis.

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Abbreviations

SVMP:

Snake venom metalloproteinase

MMPs:

Matrix metalloproteinases

SDS:

Sodium dodecylsulfate

PAGE:

Polyacrylamide gel electrophoresis

HPLC:

High performance liquid chromatography

MS:

Mass spectrometry

ACN:

Acetonitrile

TFA:

Trifluoroacetic acid

CCB:

Coomassie brilliant blue

ECM:

Extracellular matrix

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Acknowledgments

This research was supported by grants from the Science Foundation of Fujian (No. XZ04002, 2004-686).

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Authors and Affiliations

  1. Key Laboratory of Ministry of Education for Gastrointestinal Cancer, Research Center of Molecular Medicine, Fujian Medical University, Fuzhou, 350004, China

    Yi Shi, Ming-Kai Ji, Jian-Wen Xu, Xu Lin & Jian-Yin Lin

  2. Department of Molecular Pathology, Fujian Provincial Tumor Hospital, The Teaching Hospital of Fujian Medical University, Fuzhou, 350014, China

    Yi Shi

  3. Department of Dermatology, First Affiliated Hospital of Fujian Medical University, Fuzhou, 350004, China

    Ming-Kai Ji

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  1. Yi Shi
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  2. Ming-Kai Ji
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Correspondence to Jian-Yin Lin.

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Shi, Y., Ji, MK., Xu, JW. et al. High-Level Expression, Purification, Characterization and Structural Prediction of a Snake Venom Metalloproteinase Inhibitor in Pichia pastoris . Protein J 31, 212–221 (2012). https://doi.org/10.1007/s10930-012-9392-y

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