RP-HPLC binding domains of proteins

Aguilar, Marie-Isabel, Clayton, Daniel J., Holt, Phillip, Kronina, Veronica, Boysen, Reinhard I., Purcell, Anthony W. and Hearn, Milton T. W. (1998) RP-HPLC binding domains of proteins. Analytical Chemistry, 70 23: 5010-5018. doi:10.1021/ac980473c

Author Aguilar, Marie-Isabel
Clayton, Daniel J.
Holt, Phillip
Kronina, Veronica
Boysen, Reinhard I.
Purcell, Anthony W.
Hearn, Milton T. W.
Title RP-HPLC binding domains of proteins
Journal name Analytical Chemistry   Check publisher's open access policy
ISSN 0003-2700
Publication date 1998-12-01
Sub-type Article (original research)
DOI 10.1021/ac980473c
Volume 70
Issue 23
Start page 5010
End page 5018
Total pages 9
Place of publication Washington, DC, United States
Publisher American Chemical Society
Language eng
Abstract Procedures have been developed to identify the chromatographic binding domains of horse heart cytochrome c (Cyt c) and bovine growth hormone (bGH) during their interaction with reversed-phase sorbent materials. The procedure involves adsorption of the protein solute to the chromatographic sorbent, followed by proteolytic cleavage. Comparison of the proteolytic map obtained for Cyt c and bGH in free solution with the corresponding map obtained when these proteins are adsorbed to the chromatographic sorbent revealed significant differences in the digestion pattern. Following characterization of the peptides generated in both maps, the results indicated that specific regions on the surface of both Cyt c and bGH are inaccessible to tryptic cleavage when adsorbed to the hydrophobic surface of both a C-4 and a C-18 sorbent. Based on the assumption that the region of the protein surface that is in contact with the sorbent remains intact and bound to the sorbent during the digestion step, while the protein surface that is exposed to the solvent is accessible to proteolysis, the regions that were inaccessible to tryptic digestion were found to correspond to hydrophobic domains on the protein surface. These results also suggest that the three-dimensional structures of these proteins remain largely intact upon adsorption to the hydrophobic surface.
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collection: Institute for Molecular Bioscience - Publications
Version Filter Type
Citation counts: TR Web of Science Citation Count  Cited 17 times in Thomson Reuters Web of Science Article | Citations
Scopus Citation Count Cited 21 times in Scopus Article | Citations
Google Scholar Search Google Scholar
Created: Wed, 21 Mar 2012, 00:47:13 EST by Susan Allen on behalf of Institute for Molecular Bioscience