Implication of C-terminal deletion on the structure and stability of bovine beta-casein.
Bovine beta-casein (beta-CN) with its C-terminal truncated by chymosin digestion, beta-CN-(f1-192), was examined and characterized using circular dichroism (CD) under various temperature conditions. CONTIN/LL analysis of the CD data revealed significant secondary structure disruption in beta-CN-(f1-192) relative to its parent protein,beta-CN, in the temperature range (5 degrees to 70 degrees C) studied. Near-UV CD spectra indicated significant temperature dependent structural changes. Analytical ultracentrifugation results showed significant reduction but not complete abolishment of self-association in beta-CN-(f1-192) compared to whole beta-casein at 2 degrees -37 degrees C. Furthermore, binding experiments with the common hydrophobic probe - 8-anilino-1- naphthalene sulfonic acid (ANS) illustrated that beta-CN-(f1-192) is nearly incapable of binding to ANS relative to whole beta-CN, suggesting a nearly complete open overall tertiary structure brought about by the C-terminal truncation. It has been demonstrated clearly that the tail peptide beta-CN-(f193-209) is important in maintaining the hydrophobic core of beta-CN but the residual association observed argues for a minor role for other sites as well.
Qi
Phoebe X
PX
U. S. Department of Agriculture, Eastern Regional Research Center, Agricultural Research Service, 600 East Mermaid Lane, Wyndmoor, PA, 19038, USA. pqi@errc.ars.usda.gov
Wickham
Edward D
ED
Piotrowski
Edwin G
EG
Fagerquist
Clifton K
CK
Farrell
Harold M
HM
D000595:Amino Acid Sequence; D000818:Animals; D002364:Caseins / Q000737:chemistry* / Q000235:genetics; D002417:Cattle; D002942:Circular Dichroism; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D012044:Regression Analysis; D016415:Sequence Alignment; D013696:Temperature; D014461:Ultracentrifugation
D002364:Caseins