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L-arginine mediated renaturation enhances yield of human,α6 type IV collagen non-collagenous domain from bacterial inclusion bodies
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نویسنده
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gunda v. ,boosani c.s. ,verma r.k. ,guda c. ,sudhakar y.a.
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منبع
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protein and peptide letters - 2012 - دوره : 19 - شماره : 10 - صفحه:1112 -1121
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چکیده
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The anti-angiogenic,carboxy terminal non-collagenous domain (nc1) derived from human collagen type iv alpha 6 chain,[α6(iv)nc1] or hexastatin,was earlier obtained using different recombinant methods of expression in bacterial systems. however,the effect of l-arginine mediated renaturation in enhancing the relative yields of this protein from bacterial inclusion bodies has not been evaluated. in the present study,direct stirring and on-column renaturation methods using l-arginine and different size exclusion chromatography matrices were applied for enhancing the solubility in purifying the recombinant α6(iv)nc1 from bacterial inclusion bodies. this methodology enabled purification of higher quantities of soluble protein from inclusion bodies,which inhibited endothelial cell proliferation,migration and tube formation. thus,the scope for l-arginine mediated renaturation in obtaining higher yields of soluble,biologically active nc1 domain from bacterial inclusion bodies was evaluated. © 2012 bentham science publishers.
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کلیدواژه
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Anti-angiogenic activity; Human umbilical vein endothelial cells and vascular endothelial growth factor; L-arginine mediated renaturation; Non-collagenous domain of α6 type IV collagen; Size exclusion chromatography
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آدرس
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cell signaling,retinal and tumor angiogenesis laboratory,department of genetics,boys town national research hospital,omaha, United States, cell signaling,retinal and tumor angiogenesis laboratory,department of genetics,boys town national research hospital,omaha, United States, cell signaling,retinal and tumor angiogenesis laboratory,department of genetics,boys town national research hospital,omaha, United States, center for bioinformatics and systems biology,dept. of genetics,cell biology and anatomy,university of nebraska medical center,omaha, United States, cell signaling,retinal and tumor angiogenesis laboratory,department of genetics,boys town national research hospital,omaha,ne 68131,united states,department of biochemistry and molecular biology,university of nebraska medical center,omaha,ne,68198,united states,department of genetics,cell biology and anatomy,university of nebraska medical center,omaha,ne 68198,united states,department of pharmacy,school of pharmacy and health professions,creighton university,omaha, United States
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Authors
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