Abstract
The reagent o-iodosobenzoic acid (IBA)+ has been reported to cleave specifically tryptophanyl peptide bonds in proteins with 70–100% yields (1). However, the results obtained in a number of laboratories (2–4) indicated that the reagent is not as selective as originally reported, since cleavage occurs also at tyrosine in moderate to high yields. Subsequently, Mahoney et al. (5,6) proposed that o-iodoxybenzoic acid thought to be present as a contaminant in the commercially available reagent is responsible for the observed cleavage at tyrosine. The results of the studies reported herein rule out this proposal and show that IBA, under the experimental conditions of protein fragmentation (80% aqueous acetic acid containing 4M Gdn.HCl), mediates the oxidative chlorination of both the indole nucleus of tryptophan and the phenol ring of tyrosine with concomitant peptide bond cleavage++.
ArticleNote
A more comprehensive report on the fragmentation of peptides and proteins using IBA will be published elsewhere (7).
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
Abbreviations
- IBA:
-
o-iodosobenzoic acid
- BNPS-skatole:
-
2-(2-nitrophenylsulfenyl)-3-methyl-3-bromoindolenine
- NBS:
-
N-bromosuccinimide
- DMSO:
-
dimethyl sulfoxide
- Gdn.HCl:
-
guanidine hydrochloride
References
Mahoney, W.C. and Hermodson, M.A. (1979) Biochemistry 18, 3810–3814.
Fontana, A., Savige, W.A. and Zambonin, M. (1980) in Methods in Peptide and Protein Sequence Analysis (Birr, Chr., ed.) Elsevier/North–Holland Biomedical Press, Amsterdam, 309–322.
Wachter, E. and Werhahn, R. (1980) in Methods in Peptide and Protein Sequence Analysis (Birr, Chr., ed.), Elsevier/North-Holland Biomedical Press, Amsterdam, 323–328.
Johnson, P. and Stockmal, V.B. (1980) Biochem. Biophys. Res. Commun. 94, 697–703.
Mahoney, W.C. and Hermodson, M.A. (1980) in Methods in Peptide and Protein Sequence Analysis (Birr, Chr.,ed.) Elsevier North Holland Biomedical Press, Amsterdam, 323–328.
Mahoney, W.C., Smith, P.K. and Hermodson, M.A. (1981) Biochemistry 20, 443–448.
Fontana, A., Dalzoppo, D., Grandi, C. and Zambonin, M. (1981) Biochemistry, in press.
Meyer, V. and Wachter, W. (1892) Ber. 25, 2632–2635.
Askenasy, P. and Meyer, V. (1893) Ber. 26, 1354–1370.
Hellerman, L., Chinard, F.P. and Ramsdell, P.A. (1941) J. Amer. Chem. Soc. 63, 2551–2553.
Hartmann, C. and Meyer, V. (1893) Ber. 26, 1727–1732.
Spande, T.F. and Witkop, B. (1967) Methods Enzymol. 11, 498– 506.
Patchornik, A. Lawson, W.E., Gross, E. and Witkop, B. (I960) J. Amer. Chem. Soc. 82, 5923–5927.
Fontana, A. and Toniolo, C. (1976) Progr. Chem. Org. Natl. Compounds 33, 309–409.
Savige, W.E. and Fontana, A. (1980) Int. J. Peptide Protein Res. 15, 285–297.
Witkop, B. (1961) Adv. Protein Chem. 16, 221–321.
Wilson, I.G. and Cohen, L.A. (1963) J. Amer. Chem. Soc. 85, 560–564.
Ramachandran, L.K. and Witkop, B. (1967) Methods Enzymol. 11, 283–308.
Omenn, G.S., Fontana, A. and Anfinsen, C.B. (1970) J. Biol. Chem. 245, 1895–1902.
Burstein, Y. and Patchornik, A. (1972) Biochemistry 11, 4641– 4650.
Fontana, A., Vita, C. and Toniolo, C. (1973) FEBS Lett. 32, 139–142.
Savige, W.E. and Fontana, A. (1977) Methods Enzymol. 47, 459– 469.
Baker, G.P., Mann, F.G., Sheppard, N. and Tetlow, A.J. (1965) J. Chem. Soc. 3721–3728.
Fontana, A. (1972) Methods Enzymol. 25, 419–423.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1982 The HUMANA Press Inc.
About this chapter
Cite this chapter
Fontana, A., Dalzoppo, D., Grandi, C., Zambonin, M. (1982). Protein Fragmentation with o-Iodosobenzoic Acid. In: Elzinga, M. (eds) Methods in Protein Sequence Analysis. Experimental Biology and Medicine, vol 3. Humana Press. https://doi.org/10.1007/978-1-4612-5832-2_27
Download citation
DOI: https://doi.org/10.1007/978-1-4612-5832-2_27
Publisher Name: Humana Press
Print ISBN: 978-1-4612-5834-6
Online ISBN: 978-1-4612-5832-2
eBook Packages: Springer Book Archive