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. 1966 Sep;100(3):614–621. doi: 10.1042/bj1000614

Optical rotatory dispersion, circular dichroism and far-ultraviolet spectra of avidin and streptavidin

N M Green 1,*, M D Melamed 1,*
PMCID: PMC1265192  PMID: 5969276

Abstract

1. The optical-rotatory-dispersion and circular-dichroism curves of avidin showed positive Cotton effects centred at 228mμ and 280mμ, close to the ultraviolet-absorption bands of tryptophan. These effects disappeared when avidin was dissociated into sub-units in guanidine hydrochloride. 2. Binding of biotin had only a small effect on the optical-rotatory-dispersion curve of avidin. 3. The absence of negative circular dichroism at wavelengths above 216mμ showed that there was little or no α-helix present in avidin. This interpretation was confirmed by Moffitt–Yang plots of the partial rotation due to the peptide bonds in the visible region of the spectrum. The calculated dispersion constants were remarkably similar to those of γ-globulin and suggested the presence of peptide conformations other than α-helix and random coil. 4. The far-ultraviolet spectrum was also similar to that of γ-globulin, the mean extinction coefficient of the peptide chromophore being much lower than the experimental value for a random-coil structure. 5. Streptavidin resembled avidin in showing two positive Cotton effects, but the negative dichroism below 220mμ suggested the presence of more α-helix than was found in avidin. Formation of the complex with biotin was accompanied by changes in rotation that were rather larger than those observed with avidin.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BEYCHOK S., FASMAN G. D. CIRCULAR DICHROISM OF POLY-L-TYROSINE. Biochemistry. 1964 Nov;3:1675–1678. doi: 10.1021/bi00899a012. [DOI] [PubMed] [Google Scholar]
  2. Beychok S. Side-chain optical activity in cystine-containing proteins: circular dichroism studies. Proc Natl Acad Sci U S A. 1965 May;53(5):999–1006. doi: 10.1073/pnas.53.5.999. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. CHAIET L., WOLF F. J. THE PROPERTIES OF STREPTAVIDIN, A BIOTIN-BINDING PROTEIN PRODUCED BY STREPTOMYCETES. Arch Biochem Biophys. 1964 Jul 20;106:1–5. doi: 10.1016/0003-9861(64)90150-x. [DOI] [PubMed] [Google Scholar]
  4. FASMAN G. D., BODENHEIMER E., LINDBLOW C. OPTICAL ROTATORY DISPERSION STUDIES OF POLY-L-TYROSINE AND COPOLYMERS OF L-GLUTAMIC ACID AND L-TYROSINE. SIGNIFICANCE OF THE TYROSYL COTTON EFFECTS WITH RESPECT TO PROTEIN CONFORMATION. Biochemistry. 1964 Nov;3:1665–1674. doi: 10.1021/bi00899a011. [DOI] [PubMed] [Google Scholar]
  5. FUJIOKA H., IMAHORI K. Studies on the binding of carboxypeptidase A and several competitive inhibitors. J Biol Chem. 1962 Sep;237:2804–2808. [PubMed] [Google Scholar]
  6. GOULD H. J., GILL T. J., 3rd, DOTY P. THE CONFORMATION AND HYDROGEN ION EQUILIBRIUM OF NORMAL RABBIT GAMMA-GLOBULIN. J Biol Chem. 1964 Sep;239:2842–2851. [PubMed] [Google Scholar]
  7. GREEN N. M. AVIDIN. 3. THE NATURE OF THE BIOTIN-BINDING SITE. Biochem J. 1963 Dec;89:599–609. doi: 10.1042/bj0890599. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. GREEN N. M. Spectroscopic evidence for the participation of tryptophan residues in the binding of biotin by avidin. Biochim Biophys Acta. 1962 May 7;59:244–246. doi: 10.1016/0006-3002(62)90726-6. [DOI] [PubMed] [Google Scholar]
  9. HOLZWARTH G., DOTY P. THE ULTRAVIOLET CIRCULAR DICHROISM OF POLYPEPTIDES. J Am Chem Soc. 1965 Jan 20;87:218–228. doi: 10.1021/ja01080a015. [DOI] [PubMed] [Google Scholar]
  10. MARTIN N. H. SECTION OF PATHOLOGY. FORM AND FUNCTION. Proc R Soc Med. 1965 Jan;58:41–48. doi: 10.1177/003591576505800111. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. MELAMED M. D., GREEN N. M. AVIDIN. 2. PURIFICATION AND COMPOSITION. Biochem J. 1963 Dec;89:591–599. doi: 10.1042/bj0890591. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. MELAMED M. D., GREEN N. M. AVIDIN. 2. PURIFICATION AND COMPOSITION. Biochem J. 1963 Dec;89:591–599. doi: 10.1042/bj0890591. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. MYERS D. V., EDSALL J. T. OPTICAL ROTATORY DISPERSION OF HUMAN CARBONIC ANHYDRASES: COTTON EFFECTS AND AROMATIC ABSORPTION BANDS. Proc Natl Acad Sci U S A. 1965 Jan;53:169–177. doi: 10.1073/pnas.53.1.169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. ROSENHECK K., DOTY P. The far ultraviolet absorption spectra of polypeptide and protein solutions and their dependence on conformation. Proc Natl Acad Sci U S A. 1961 Nov 15;47:1775–1785. doi: 10.1073/pnas.47.11.1775. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Sarkar P. K., Doty P. The optical rotatory properties of the beta-configuration in polypeptides and proteins. Proc Natl Acad Sci U S A. 1966 Apr;55(4):981–989. doi: 10.1073/pnas.55.4.981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Simmons N. S., Blout E. R. The Structure of Tobacco Mosaic Virus and Its Components: Ultraviolet Optical Rotatory Dispersion. Biophys J. 1960 Sep;1(1):55–62. doi: 10.1016/s0006-3495(60)86875-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. URRY D. W., DOTY P. ON THE CONFORMATION OF HORSE HEART FERRI- AND FERROCYTOCHROME C. J Am Chem Soc. 1965 Jun 20;87:2756–2758. doi: 10.1021/ja01090a041. [DOI] [PubMed] [Google Scholar]
  18. Ulmer D. D., Vallee B. L. Extrinsic cotton effects and the mechanism of enzyme action. Adv Enzymol Relat Areas Mol Biol. 1965;27:37–104. doi: 10.1002/9780470122723.ch2. [DOI] [PubMed] [Google Scholar]
  19. Van Holde K. E., Brahms J., Michelson A. M. Base interactions of nucleotide polymers in aqueous solution. J Mol Biol. 1965 Jul;12(3):726–739. doi: 10.1016/s0022-2836(65)80323-0. [DOI] [PubMed] [Google Scholar]

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