L-arginine mediated renaturation enhances yield of human, α6 type IV collagen non-collagenous domain from bacterial inclusion bodies

Venugopal Gunda, Chandra Shekhar Boosani, Raj Kumar Verma, Chittibabu Guda, Yakkanti Akul Sudhakar

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)1112-1121
Number of pages10
JournalProtein and Peptide Letters
Volume19
Issue number10
DOIs
StatePublished - Oct 1 2012

Fingerprint

Collagen Type VI
Collagen Type IV
Inclusion Bodies
Arginine
Bacterial Proteins
Size exclusion chromatography
Endothelial cells
Cell proliferation
Solubility
Cell Movement
Purification
Gel Chromatography
Endothelial Cells
Cell Proliferation
Proteins

Keywords

  • 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

ASJC Scopus subject areas

  • Structural Biology
  • Biochemistry

Cite this

L-arginine mediated renaturation enhances yield of human, α6 type IV collagen non-collagenous domain from bacterial inclusion bodies. / Gunda, Venugopal; Boosani, Chandra Shekhar; Verma, Raj Kumar; Guda, Chittibabu; Sudhakar, Yakkanti Akul.

In: Protein and Peptide Letters, Vol. 19, No. 10, 01.10.2012, p. 1112-1121.

Research output: Contribution to journalArticle

Gunda, Venugopal ; Boosani, Chandra Shekhar ; Verma, Raj Kumar ; Guda, Chittibabu ; Sudhakar, Yakkanti Akul. / L-arginine mediated renaturation enhances yield of human, α6 type IV collagen non-collagenous domain from bacterial inclusion bodies. In: Protein and Peptide Letters. 2012 ; Vol. 19, No. 10. pp. 1112-1121.
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