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Off-pathway status for the alkali molten globule of horse ferricytochrome C.
Biochemistry. 2010 Sep 14; 49(36):7764-73.B

Abstract

The Na(+)-stabilized alkali molten globule (B-state) of horse ferricytochrome c produced at pH 12.9 in 2 M NaCl has been studied to find out its relevance to the kinetic folding pathway of the protein. Details of stopped-flow kinetics indicate that the B-state when driven to fold at pH 11.5 goes through a submillisecond burst expansion to a state B' which must unfold to the base-denatured U(B)-state before folding to the alkaline native state, N(B). This folding hallmark suggests that the B-state is a dead-end or off-pathway species, playing apparently an unclear role in the folding kinetics. Interestingly, the folding kinetics of the B-state at a final pH of 7 is very similar to that observed for the guanidinium-unfolded ferricytochrome c (U) at pH 7. Both B and U exhibit a submillisecond burst phase followed by three observable phases, fast, medium, and slow, with matching rate constants for the fast phase, suggesting that B- and U-states share a common folding mechanism. Even the minima in the folding chevrons for the B- and U-states appear at the same denaturant concentration, but the former is shifted vertically upward and has shallower limbs, suggesting that the transition state relevant for the B-state folding is relatively more compact with greater surface burial allaying large-scale diffusive migration of chain segments. It is concluded that the B-state is not a good model for the kinetic molten globule of cytochrome c. Part of the reason for such atypical response of a typical molten globule may possibly be related to misligation of the ferric heme with lysyl side chains at the extreme alkaline pH required to produce the B-state. To eliminate this possibility, the companion paper [Bhuyan, A. K. (2010) Biochemistry (DOI 10.1021/bi100881n)] studies the B-state of ferrocytochrome c where ligation of the heme with any intrapolypeptide side chain is completely suppressed. The study concludes that the B-state is kinetically an abortive species.

Authors+Show Affiliations

School of Chemistry, University of Hyderabad, Hyderabad 500046, India. akbsc@uohyd.ernet.in

Pub Type(s)

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

Language

eng

PubMed ID

20687523

Citation

Bhuyan, Abani K.. "Off-pathway Status for the Alkali Molten Globule of Horse Ferricytochrome C." Biochemistry, vol. 49, no. 36, 2010, pp. 7764-73.
Bhuyan AK. Off-pathway status for the alkali molten globule of horse ferricytochrome C. Biochemistry. 2010;49(36):7764-73.
Bhuyan, A. K. (2010). Off-pathway status for the alkali molten globule of horse ferricytochrome C. Biochemistry, 49(36), 7764-73. https://doi.org/10.1021/bi100880d
Bhuyan AK. Off-pathway Status for the Alkali Molten Globule of Horse Ferricytochrome C. Biochemistry. 2010 Sep 14;49(36):7764-73. PubMed PMID: 20687523.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Off-pathway status for the alkali molten globule of horse ferricytochrome C. A1 - Bhuyan,Abani K, PY - 2010/8/7/entrez PY - 2010/8/7/pubmed PY - 2010/11/4/medline SP - 7764 EP - 73 JF - Biochemistry JO - Biochemistry VL - 49 IS - 36 N2 - The Na(+)-stabilized alkali molten globule (B-state) of horse ferricytochrome c produced at pH 12.9 in 2 M NaCl has been studied to find out its relevance to the kinetic folding pathway of the protein. Details of stopped-flow kinetics indicate that the B-state when driven to fold at pH 11.5 goes through a submillisecond burst expansion to a state B' which must unfold to the base-denatured U(B)-state before folding to the alkaline native state, N(B). This folding hallmark suggests that the B-state is a dead-end or off-pathway species, playing apparently an unclear role in the folding kinetics. Interestingly, the folding kinetics of the B-state at a final pH of 7 is very similar to that observed for the guanidinium-unfolded ferricytochrome c (U) at pH 7. Both B and U exhibit a submillisecond burst phase followed by three observable phases, fast, medium, and slow, with matching rate constants for the fast phase, suggesting that B- and U-states share a common folding mechanism. Even the minima in the folding chevrons for the B- and U-states appear at the same denaturant concentration, but the former is shifted vertically upward and has shallower limbs, suggesting that the transition state relevant for the B-state folding is relatively more compact with greater surface burial allaying large-scale diffusive migration of chain segments. It is concluded that the B-state is not a good model for the kinetic molten globule of cytochrome c. Part of the reason for such atypical response of a typical molten globule may possibly be related to misligation of the ferric heme with lysyl side chains at the extreme alkaline pH required to produce the B-state. To eliminate this possibility, the companion paper [Bhuyan, A. K. (2010) Biochemistry (DOI 10.1021/bi100881n)] studies the B-state of ferrocytochrome c where ligation of the heme with any intrapolypeptide side chain is completely suppressed. The study concludes that the B-state is kinetically an abortive species. SN - 1520-4995 UR - https://www.unboundmedicine.com/medline/citation/20687523/Off_pathway_status_for_the_alkali_molten_globule_of_horse_ferricytochrome_C_ L2 - https://doi.org/10.1021/bi100880d DB - PRIME DP - Unbound Medicine ER -