Abstract
Bovine β-casein (β-CN) with its C-terminal truncated by chymosin digestion, β-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 β-CN-(f1-192) relative to its parent protein,β-CN, in the temperature range (5° to 70°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 β-CN-(f1-192) compared to whole β-casein at 2°–37°C. Furthermore, binding experiments with the common hydrophobic probe – 8-anilino-1- naphthalene sulfonic acid (ANS) illustrated that β-CN-(f1-192) is nearly incapable of binding to ANS relative to whole β-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 β-CN-(f193-209) is important in maintaining the hydrophobic core of β-CN but the residual association observed argues for a minor role for other sites as well.
Similar content being viewed by others
Abbreviations
- ACS:
-
American Chemical Society
- ANS:
-
1-Anilino-8-Naphthalene Sulfonic Acid
- CD:
-
Circular Dichroism
- ESI-MS:
-
Electronspray Ionization Mass Spectrometry
- FTIR:
-
Fourier Transform Infrared Spectroscopy
- MG:
-
Molten Globule
- MW:
-
Molecular Weight
- ORD:
-
Optical Rotatory Dispersion
- PIPES:
-
Piperazine-N,N´-bis(2-Ethanesulfonic Acid)
- SDS:
-
Sodium Dodecyl Sulfate
- PPII:
-
Left handed polyproline II helix
- r.m.s.d.:
-
Root Mean Square Deviation
- UV:
-
Ultra Violet.
References
A. L. Andrews D. Atkinson M. T. A. Evans E. G. Finer J. P. Green M. C. Phillips N. Robertson (1979) Biopolymers 18 1105–1121 Occurrence Handle1:CAS:528:DyaE1MXktVWlsrc%3D Occurrence Handle10.1002/bip.1979.360180507
G. P. Berry L. K. Creamer (1975) Biochemistry 14 3542–3545 Occurrence Handle1:CAS:528:DyaE2MXlt1Okt7g%3D Occurrence Handle10.1021/bi00687a005
H. Bu S. M. Sood C. W. Slattery (2004) Protein J. 23 509–517 Occurrence Handle1:CAS:528:DC%2BD2cXhtVeju7rO Occurrence Handle10.1007/s10930-004-7878-y
L. K. Creamer O. E. Mills E. L. Richards (1971) J. Dairy Res. 38 269–280 Occurrence Handle1:CAS:528:DyaE38XlsF2ntw%3D%3D
S. Damodaran L. Rammovsky (2003) Food Hydrocolloids 17 355–363 Occurrence Handle1:CAS:528:DC%2BD3sXjsFyns70%3D Occurrence Handle10.1016/S0268-005X(02)00098-X
W. L. DeLano M. H. Ultsch A. M. Vos Particlede J. A. Wells (2000) Science 287 1279–1283 Occurrence Handle1:CAS:528:DC%2BD3cXhtlOqsbg%3D Occurrence Handle10.1126/science.287.5456.1279
H. J. Dyson P. E. Wright (2005) Nat. Rev. Mol. Cell Biol. 6 197–208 Occurrence Handle1:CAS:528:DC%2BD2MXhslSlsLo%3D Occurrence Handle10.1038/nrm1589
H. M. Farrell SuffixJr. (1999) NoChapterTitle E. Knobil J. D. Neill (Eds) Encyclopedia of Reproduction NumberInSeriesVol. III Academic Press, Inc., San Diego CA 256–263
H. M. Farrell SuffixJr. J. T. Deeney E. K. Hild T. F. Kumosinski (1990) J. Biol. Chem. 265 17637–17643 Occurrence Handle1:CAS:528:DyaK3cXmtVOgsL4%3D
H. M. Farrell SuffixJr. R. Jimenez-Flores G. T. Bleck E. M. Brown J. E. Butler L. K. Creamer C. L. Hicks C. M. Hollar K. F. Ng-Kwai-Hang H. E. Swaisgood (2004) J. Dairy Sci. 87 1641–1674 Occurrence Handle1:CAS:528:DC%2BD2cXltlGnsrk%3D
H. M. Farrell SuffixJr. T. F. Kumosinski P. Pulaski M. P. Thompson (1988) Arch. Biochem. Biophys 265 146–158 Occurrence Handle1:CAS:528:DyaL1cXmtVKqtr0%3D Occurrence Handle10.1016/0003-9861(88)90380-3
H. M. Farrell SuffixJr. P. X. Qi (2005) NoChapterTitle M. L. Fishman P. X. Qi L. Wicker (Eds) Advances in Biopolymers: Molecules, clusters, networks and interactions American Chemical Society Washington, D.C
H. M. Farrell SuffixJr. P. X. Qi E. D. Wickham J. J. Unruh (2002) J. Protein Chem. 21 307–321 Occurrence Handle1:CAS:528:DC%2BD38XmsVGlsrk%3D
H. M. Farrell SuffixJr. E. D. Wickham J. J. Unruh P. X. Qi P. D. Hoagland (2001) Food Hydrocolloids 15 341–354 Occurrence Handle1:CAS:528:DC%2BD3MXosVCksbw%3D Occurrence Handle10.1016/S0268-005X(01)00080-7
G. Fimland V. G. H. Eijsink J. Nissen-Meyer (2002) Biochemistry 41 9508–9515 Occurrence Handle1:CAS:528:DC%2BD38XkvVygtrg%3D Occurrence Handle10.1021/bi025856q
J. Garnier (1966) J. Mol. Biol. 19 586–590 Occurrence Handle1:STN:280:DyaF2s7ntlOmug%3D%3D Occurrence Handle10.1016/S0022-2836(66)80028-1
E. R. B. Graham G. N. Malcolm H. A. McKenzie (1984) Int. J. Biol. Macromol. 6 155–161 Occurrence Handle1:CAS:528:DyaL2cXks1yjsb0%3D Occurrence Handle10.1016/0141-8130(84)90058-8
T. T. Herskovits (1966) Biochemistry 5 1018–1026 Occurrence Handle1:CAS:528:DyaF28XmvFWrtQ%3D%3D Occurrence Handle10.1021/bi00867a030
C. Holt L. Sawyer (1993) J. Chem. Soc. Faraday Trans. 89 2683–2692 Occurrence Handle1:CAS:528:DyaK3sXmt1Sjsr4%3D Occurrence Handle10.1039/ft9938902683
D. Matulis R. Lovrien (1998) Biophys. J. 74 422–429 Occurrence Handle1:CAS:528:DyaK1cXhvFSqurk%3D Occurrence Handle10.1016/S0006-3495(98)77799-9
M. Noelken M. Reibstein (1968) Arch. Biochem. Biophys. 123 397–402 Occurrence Handle1:CAS:528:DyaF1cXls1KjsQ%3D%3D Occurrence Handle10.1016/0003-9861(68)90150-1
M. Pagani S. Pilati G. Bertoli B. Valsasina R. Sitia (2001) FEBS Lett. 508 117–120 Occurrence Handle1:CAS:528:DC%2BD3MXot1OmtL8%3D Occurrence Handle10.1016/S0014-5793(01)03034-4
S. Patton (2004) Milk: Its remarkable contribution to human health and well-being Transaction Publishers New Brunswick, NJ
T. A. J. Payens B. W. Markwijk Particlevan (1963) Biochim. Biophys. Acta. 71 517–530 Occurrence Handle1:CAS:528:DyaF3sXktlKlsr4%3D Occurrence Handle10.1016/0006-3002(63)91124-7
S. W. Provencher J. Glockner (1981) Biochemstry 20 33–37 Occurrence Handle1:CAS:528:DyaL3MXis12nsg%3D%3D
P. X. Qi (2005) NoChapterTitle M. L. Fishman P. X. Qi L. Wicker (Eds) Advances in Biopolymers: Molecules, clusters, networks and interactions American Chemical Society Washington, D.C
P. X. Qi E. D. Wickham H. M. Farrell SuffixJr. (2004) Protein J. 23 389–402 Occurrence Handle1:CAS:528:DC%2BD2cXntVKqsrg%3D Occurrence Handle10.1023/B:JOPC.0000039553.66233.3f
J. S. Rodriguez-Zavala H. Weiner (2002) Biochemistry 41 8229–8237 Occurrence Handle1:CAS:528:DC%2BD38Xkt1aisb0%3D Occurrence Handle10.1021/bi012081x
A. L. Rucker T. P. Creamer (2002) Protein Sci. 11 980–985 Occurrence Handle1:CAS:528:DC%2BD38Xis1Wls7w%3D
J. F. Rusling T. F. Kumosinski (1996) Nonlinear computer modeling of chemical and biochemical data Academic Press San Diego, CA
V. P. Saxena D. B. Wetlaufer (1971) Proc. Natl. Acad. Sci. USA 68 969–972 Occurrence Handle1:CAS:528:DyaE3MXktlKltbg%3D
D. G. Schmidt (1979) J. Dairy Res. 46 351–355 Occurrence Handle1:CAS:528:DyaE1MXltlGkurk%3D
D. G. Schmidt (1982) NoChapterTitle P. F. Fox (Eds) Developments in Dairy Chemistry NumberInSeriesVol. I Applied Science London 61–86
D. G. Schmidt T. A. J. Payens (1976) NoChapterTitle E. Matijevic (Eds) Surface and Colloid Science NumberInSeriesVol. IX John Wiley, New York NY 165–199
G. V. Semisotnov N. A. Rodionova O. I. Razgulyaev V. N. Uversky A. F. Gripas R. I. Gilmanshin (1991) Biopolymers 31 119–128 Occurrence Handle1:CAS:528:DyaK3MXhvVWmurY%3D Occurrence Handle10.1002/bip.360310111
Z. Shi R. W. Woody N. R. Kallenbach (2002) Adv. Protein Chem. 62 163–240 Occurrence Handle1:CAS:528:DC%2BD38XpsVeqs7s%3D Occurrence Handle10.1016/S0065-3233(02)62008-X
N. Sreerama S. Y. Venyaminov R. W. Woody (2000) Anal. Biochem. 287 243–251 Occurrence Handle1:CAS:528:DC%2BD3cXosFGhu7k%3D
N. Sreerama R. W. Woody (2000) Anal. Biochem. 287 252–260 Occurrence Handle1:CAS:528:DC%2BD3cXosFGhu7Y%3D
C. D. Syme E. W. Blanch C. Holt R. Jakes M. Goedert L. Hecht L. D. Barron (2002) Eur. J. Biochem. 269 148–156 Occurrence Handle1:CAS:528:DC%2BD38XosFGisw%3D%3D Occurrence Handle10.1046/j.0014-2956.2001.02633.x
M. Tai G. Kegeles (1984) Biophys. Chem. 20 81–87 Occurrence Handle1:CAS:528:DyaL2cXlsVKgu7s%3D Occurrence Handle10.1016/0301-4622(84)80007-1
K. Takase R. Niki S. Arima (1980) Biochim. Biophys. Acta 622 1–8 Occurrence Handle1:CAS:528:DyaL3cXhvFejsr4%3D
M. P. Thompson (1966) J. Dairy Sci. 49 792–795 Occurrence Handle1:CAS:528:DyaF28XktlWgsbY%3D
M. P. Thompson E. B. Kalan R. Greenberg (1967) J. Dairy Sci. 50 767–769 Occurrence Handle1:CAS:528:DyaF2sXksVShsrs%3D Occurrence Handle10.3168/jds.S0022-0302(67)87511-8
A. Thurn W. Burchard R. Niki (1987) Polymer Sci. 265 653–666 Occurrence Handle1:CAS:528:DyaL2sXlvFKksr8%3D
M. L. Tiffany S. Krimm (1968) Biopolymers 6 1379–1382 Occurrence Handle1:CAS:528:DyaF1cXkslOrur4%3D
P. Tompa (2002) Trends Biochem. Sci. 27 527–533 Occurrence Handle1:CAS:528:DC%2BD38XnsVWqsbo%3D Occurrence Handle10.1016/S0968-0004(02)02169-2
A. Toumadje W. C. Johnson SuffixJr. (1995) J. Am. Chem. Soc. 117 7023–7024 Occurrence Handle1:CAS:528:DyaK2MXmsFSmt7g%3D Occurrence Handle10.1021/ja00131a034
V. N. Uversky (2002a) Eur. J. Biochem 269 2–12 Occurrence Handle1:CAS:528:DC%2BD38XosFCqtg%3D%3D Occurrence Handle10.1046/j.0014-2956.2001.02649.x
V. N. Uversky (2002b) Protein Sci 11 739–756 Occurrence Handle1:CAS:528:DC%2BD38Xis1Wmu74%3D Occurrence Handle10.1110/ps.4210102
B. Valeur G. Weber (1977) Photochem. Photobiol. 25 441–444 Occurrence Handle1:CAS:528:DyaE2sXltValsLg%3D
S. Visser K. J. Slangen (1977) Neth. Milk Dairy J. 31 16–30 Occurrence Handle1:CAS:528:DyaE2sXktlCqt74%3D
N. M. Wahlgren P. Dejmek T. Drakenberg (1994) J. Dairy Res. 61 495–506 Occurrence Handle1:CAS:528:DyaK2MXit1Gktbk%3D Occurrence Handle10.1017/S0022029900028429
N. M. Wahlgren J. Leonil P. Dejmek T. Drakenberg (1993) Biochim. Biophys. Acta 1202 121–128 Occurrence Handle1:CAS:528:DyaK2cXpvFOhsw%3D%3D
D. F. Waugh L. K. Creamer C. W. Slattery G. W. Dresdner (1970) Biochemistry 9 786–795 Occurrence Handle1:CAS:528:DyaE3cXps1Krtw%3D%3D Occurrence Handle10.1021/bi00806a011
D. B. Wetlaufer (1962) Adv. Protein Chem. 17 303–391 Occurrence Handle1:CAS:528:DyaF3sXktV2qsb0%3D
J. Wyman SuffixJr. (1964) Adv. Protein Chem. 19 223–286 Occurrence Handle1:CAS:528:DyaF2MXitlCnuw%3D%3D
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Qi, P.X., Wickham, E.D., Piotrowski, E.G. et al. Implication of C-Terminal Deletion on the Structure and Stability of Bovine β-casein. Protein J 24, 431–444 (2005). https://doi.org/10.1007/s10930-005-7639-6
Issue Date:
DOI: https://doi.org/10.1007/s10930-005-7639-6