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Native-like beta-hairpin retained in the cold-denatured state of bovine beta-lactoglobulin.
J Mol Biol. 2001 Jul 06; 310(2):471-84.JM

Abstract

Bovine beta-lactoglobulin is denatured by increased temperature (heat denaturation) and by decreased temperature (cold-denaturation) in the presence of 4 M urea at pH 2.5. We characterized the structure of the cold-denatured state of beta-lactoglobulin using circular dichroism (CD), small-angle X-ray scattering (SAXS) and heteronuclear nuclear magnetic resonance (NMR). CD and SAXS indicated that the cold-denatured state, in comparison with the highly denatured state induced by urea, is rather compact, retaining some secondary structure, but no tertiary structure. The location of the residual structures in the cold-denatured state and their stability were characterized by 1H/2H exchange combined with heteronuclear NMR. The results indicated that the residues adjacent to the disulfide bond (C106-C119) connecting beta-strands G and H had markedly high protection factors, suggesting the presence of a native-like beta-hairpin stabilized by the disulfide bond. Since this beta-hairpin is conserved between different conformational states, including the kinetic refolding intermediate, it should be of paramount importance for the folding and stability of beta-lactoglobulin. On the other hand, the non-native alpha-helix suggested for the folding intermediate was not detected in the cold-denatured state. The 1H/2H exchange experiments showed that the protection factors of a mixture of the native and cold-denatured states is strongly biased by that of the labile cold-denatured state, consistent with a two-process model of the exchange.

Authors+Show Affiliations

Institute for Protein Research, Osaka University, Yamadaoka 3-2, Suita, Osaka, 565-0871, Japan.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

11428901

Citation

Katou, H, et al. "Native-like Beta-hairpin Retained in the Cold-denatured State of Bovine Beta-lactoglobulin." Journal of Molecular Biology, vol. 310, no. 2, 2001, pp. 471-84.
Katou H, Hoshino M, Kamikubo H, et al. Native-like beta-hairpin retained in the cold-denatured state of bovine beta-lactoglobulin. J Mol Biol. 2001;310(2):471-84.
Katou, H., Hoshino, M., Kamikubo, H., Batt, C. A., & Goto, Y. (2001). Native-like beta-hairpin retained in the cold-denatured state of bovine beta-lactoglobulin. Journal of Molecular Biology, 310(2), 471-84.
Katou H, et al. Native-like Beta-hairpin Retained in the Cold-denatured State of Bovine Beta-lactoglobulin. J Mol Biol. 2001 Jul 6;310(2):471-84. PubMed PMID: 11428901.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Native-like beta-hairpin retained in the cold-denatured state of bovine beta-lactoglobulin. AU - Katou,H, AU - Hoshino,M, AU - Kamikubo,H, AU - Batt,C A, AU - Goto,Y, PY - 2001/6/29/pubmed PY - 2001/7/28/medline PY - 2001/6/29/entrez SP - 471 EP - 84 JF - Journal of molecular biology JO - J Mol Biol VL - 310 IS - 2 N2 - Bovine beta-lactoglobulin is denatured by increased temperature (heat denaturation) and by decreased temperature (cold-denaturation) in the presence of 4 M urea at pH 2.5. We characterized the structure of the cold-denatured state of beta-lactoglobulin using circular dichroism (CD), small-angle X-ray scattering (SAXS) and heteronuclear nuclear magnetic resonance (NMR). CD and SAXS indicated that the cold-denatured state, in comparison with the highly denatured state induced by urea, is rather compact, retaining some secondary structure, but no tertiary structure. The location of the residual structures in the cold-denatured state and their stability were characterized by 1H/2H exchange combined with heteronuclear NMR. The results indicated that the residues adjacent to the disulfide bond (C106-C119) connecting beta-strands G and H had markedly high protection factors, suggesting the presence of a native-like beta-hairpin stabilized by the disulfide bond. Since this beta-hairpin is conserved between different conformational states, including the kinetic refolding intermediate, it should be of paramount importance for the folding and stability of beta-lactoglobulin. On the other hand, the non-native alpha-helix suggested for the folding intermediate was not detected in the cold-denatured state. The 1H/2H exchange experiments showed that the protection factors of a mixture of the native and cold-denatured states is strongly biased by that of the labile cold-denatured state, consistent with a two-process model of the exchange. SN - 0022-2836 UR - https://www.unboundmedicine.com/medline/citation/11428901/Native_like_beta_hairpin_retained_in_the_cold_denatured_state_of_bovine_beta_lactoglobulin_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0022-2836(01)94777-1 DB - PRIME DP - Unbound Medicine ER -