Changes in the secondary structure of bovine casein by Fourier transform infrared spectroscopy: effects of calcium and temperature.
Bovine casein submicelles and reformed micelles, produced by addition of Ca2+, were examined by Fourier transform infrared spectroscopy at 15 and 37 degrees C in aqueous salt solutions of K+ and Na+. Previous measurements of caseins, made in D2O and in the solid form, can now be made in a more realistic environment of H2O. When analyzed in detail, data obtained by Fourier transform infrared spectroscopy have the potential to show subtle changes in secondary structural elements that are associated with changes in protein environment. Electrostatic binding of Ca2+ to casein resulted in a redistribution of the components of the infrared spectra. Addition of Ca2+ in salt solutions of K+ and Na+ led to apparent decreases in large loop or helical structures at 37 degrees C with concomitant increases in the percentage of structures having greater bond energy, such as turns and extended helical structures. At 15 degrees C, Na+ and K+ have differential effects on the Ca(2+)-casein complexes. All of these observations are in accordance with the important role of serine phosphate side chains as sites for Ca2+ binding in caseins and the swelling of the casein structure upon incorporation into reformed micelles at 37 degrees C. This new open, hydrated structure is buttressed by a change in backbone as evidenced by a shift in absorbance to higher wave numbers (greater bond energies) as colloidal micelles are reformed.
Curley
D M
DM
USDA, Agricultural Research Service, Eastern Regional Research Center, Wyndmoor, PA 19038, USA.
Kumosinski
T F
TF
Unruh
J J
JJ
Farrell
H M
HM
D000818:Animals; D002118:Calcium / Q000378:metabolism / Q000494:pharmacology*; D002364:Caseins / Q000737:chemistry* / Q000378:metabolism; D002417:Cattle; D005260:Female; D011188:Potassium / Q000494:pharmacology; D017433:Protein Structure, Secondary; D012964:Sodium / Q000494:pharmacology; D017550:Spectroscopy, Fourier Transform Infrared; D055672:Static Electricity; D013696:Temperature
D002364:Caseins; D012964:Sodium; D011188:Potassium; D002118:Calcium