Purification and renaturation of membrane neuraminidase from Haemophilus parasuis
| Autor: | Carol A. Lichtensteiger, Eric R. Vimr |
|---|---|
| Rok vydání: | 2003 |
| Předmět: |
Protein Denaturation
Swine Protein Renaturation Haemophilus Neuraminidase Virulence Sialidase Microbiology law.invention law Protein purification Animals chemistry.chemical_classification General Veterinary biology Molecular mass General Medicine Chromatography Ion Exchange biology.organism_classification Virology Molecular Weight Enzyme chemistry Recombinant DNA biology.protein Electrophoresis Polyacrylamide Gel Bacterial Outer Membrane Proteins |
| Zdroj: | Veterinary Microbiology. 93:79-87 |
| ISSN: | 0378-1135 |
| DOI: | 10.1016/s0378-1135(02)00443-1 |
| Popis: | Haemophilus parasuis, which causes polyserositis, polysynovitis, meningitis, septicemia, and pneumonia in pigs, has emerged as an increasing problem in modern swine production systems. Co-factors for and the pathogenesis of H. parasuis disease are not defined. One of the potential virulence factors of H. parasuis is its neuraminidase (sialidase). While purifying the H. parasuis neuraminidase from the membrane fraction, we developed a protocol to renature enzymatic activity after enzyme preparations were resolved electrophorectically in denaturing polyacrylamide gels. The H. parasuis neuraminidase co-resolved with recombinant neuraminidase of Vibrio cholera; thus its apparent molecular mass is 82 kilodalton (kDa). The H. parasuis neuraminidase was associated with the membrane fraction and the purification protocol removed over 99% of the H. parasuis cell protein while retaining over 90% of the neuraminidase activity. Purified protein will provide another avenue to clone the neuraminidase gene that has been refractory to cloning and the protocol will be a means to purify recombinant protein. |
| Databáze: | OpenAIRE |
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