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Structural, kinetic and thermodynamic characterizations of SDS-induced molten globule state of a highly negatively charged cytochrome c
Abstract This study presents the structural, kinetic and thermodynamic characterizations of previously unknown submicellar concentrations of SDS-induced molten globule (MGSDS) state of a highly negatively charged base-denatured ferricytochrome c (UB-state) at pH ∼12.8 (±0.2). The far-UV CD, near-UV...
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Published in: | Journal of biochemistry (Tokyo) 2019-02, Vol.165 (2), p.125-137 |
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container_title | Journal of biochemistry (Tokyo) |
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creator | Jain, Rishu Sharma, Deepak Kumar, Rakesh Kumar, Rajesh |
description | Abstract
This study presents the structural, kinetic and thermodynamic characterizations of previously unknown submicellar concentrations of SDS-induced molten globule (MGSDS) state of a highly negatively charged base-denatured ferricytochrome c (UB-state) at pH ∼12.8 (±0.2). The far-UV CD, near-UV CD, ANS-fluorescence data of UB-state in the presence of different concentrations of SDS indicate that the submicellar concentrations of SDS (≤0.4 mM) transform the UB-state to MGSDS-state. The MGSDS-state has native-like α-helical secondary structure but lacks tertiary structure. The free energy change (ΔG°D) for UB→ MGSDS transition determined by far-UV CD (∼2.7 kcal mol−1) is slightly higher than those determined by fluorescence (∼2.0 kcal mol−1) at 25°C. At very low SDS and NaCl concentrations, the MGSDS-state undergoes cold denaturation. As SDS concentration is increased, the thermal denaturation temperature increases and the cold denaturation temperature decrease. Kinetic experiments involving the measurement of the CO-association rate to the base-denatured ferrocytochrome c at pH ≈12.8 (±0.2), 25°C indicate that the submicellar concentrations of SDS restrict the internal dynamics of base-denatured protein. |
doi_str_mv | 10.1093/jb/mvy087 |
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This study presents the structural, kinetic and thermodynamic characterizations of previously unknown submicellar concentrations of SDS-induced molten globule (MGSDS) state of a highly negatively charged base-denatured ferricytochrome c (UB-state) at pH ∼12.8 (±0.2). The far-UV CD, near-UV CD, ANS-fluorescence data of UB-state in the presence of different concentrations of SDS indicate that the submicellar concentrations of SDS (≤0.4 mM) transform the UB-state to MGSDS-state. The MGSDS-state has native-like α-helical secondary structure but lacks tertiary structure. The free energy change (ΔG°D) for UB→ MGSDS transition determined by far-UV CD (∼2.7 kcal mol−1) is slightly higher than those determined by fluorescence (∼2.0 kcal mol−1) at 25°C. At very low SDS and NaCl concentrations, the MGSDS-state undergoes cold denaturation. As SDS concentration is increased, the thermal denaturation temperature increases and the cold denaturation temperature decrease. Kinetic experiments involving the measurement of the CO-association rate to the base-denatured ferrocytochrome c at pH ≈12.8 (±0.2), 25°C indicate that the submicellar concentrations of SDS restrict the internal dynamics of base-denatured protein.</description><identifier>ISSN: 0021-924X</identifier><identifier>EISSN: 1756-2651</identifier><identifier>DOI: 10.1093/jb/mvy087</identifier><identifier>PMID: 30371870</identifier><language>eng</language><publisher>England: Oxford University Press</publisher><subject>Cytochromes c - chemistry ; Cytochromes c - metabolism ; Kinetics ; Protein Denaturation ; Sodium Chloride - chemistry ; Sodium Dodecyl Sulfate - chemistry ; Thermodynamics</subject><ispartof>Journal of biochemistry (Tokyo), 2019-02, Vol.165 (2), p.125-137</ispartof><rights>The Author(s) 2018. Published by Oxford University Press on behalf of the Japanese Biochemical Society. All rights reserved 2018</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c365t-c724e82c0b4258d89c22999f0f496080243917311737bd670fd234171dc1e0873</citedby><cites>FETCH-LOGICAL-c365t-c724e82c0b4258d89c22999f0f496080243917311737bd670fd234171dc1e0873</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/30371870$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Jain, Rishu</creatorcontrib><creatorcontrib>Sharma, Deepak</creatorcontrib><creatorcontrib>Kumar, Rakesh</creatorcontrib><creatorcontrib>Kumar, Rajesh</creatorcontrib><title>Structural, kinetic and thermodynamic characterizations of SDS-induced molten globule state of a highly negatively charged cytochrome c</title><title>Journal of biochemistry (Tokyo)</title><addtitle>J Biochem</addtitle><description>Abstract
This study presents the structural, kinetic and thermodynamic characterizations of previously unknown submicellar concentrations of SDS-induced molten globule (MGSDS) state of a highly negatively charged base-denatured ferricytochrome c (UB-state) at pH ∼12.8 (±0.2). The far-UV CD, near-UV CD, ANS-fluorescence data of UB-state in the presence of different concentrations of SDS indicate that the submicellar concentrations of SDS (≤0.4 mM) transform the UB-state to MGSDS-state. The MGSDS-state has native-like α-helical secondary structure but lacks tertiary structure. The free energy change (ΔG°D) for UB→ MGSDS transition determined by far-UV CD (∼2.7 kcal mol−1) is slightly higher than those determined by fluorescence (∼2.0 kcal mol−1) at 25°C. At very low SDS and NaCl concentrations, the MGSDS-state undergoes cold denaturation. As SDS concentration is increased, the thermal denaturation temperature increases and the cold denaturation temperature decrease. Kinetic experiments involving the measurement of the CO-association rate to the base-denatured ferrocytochrome c at pH ≈12.8 (±0.2), 25°C indicate that the submicellar concentrations of SDS restrict the internal dynamics of base-denatured protein.</description><subject>Cytochromes c - chemistry</subject><subject>Cytochromes c - metabolism</subject><subject>Kinetics</subject><subject>Protein Denaturation</subject><subject>Sodium Chloride - chemistry</subject><subject>Sodium Dodecyl Sulfate - chemistry</subject><subject>Thermodynamics</subject><issn>0021-924X</issn><issn>1756-2651</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2019</creationdate><recordtype>article</recordtype><recordid>eNp9kMtKxDAUhoMoOl4WvoBk4ULBai6dplmKdxBcjIK7kian045tMyapUF_A1zbDqEsXh3PhO__iQ-iQknNKJL9YlBfdx0hysYEmVEyzhGVTuokmhDCaSJa-7qBd7xerlXG-jXY44YLmgkzQ1yy4QYfBqfYMvzU9hEZj1RscanCdNWOvunjRtXJKB3DNpwqN7T22FZ5dz5KmN4MGgzvbBujxvLXl0AL2QQVYMQrXzbxuR9zDPH5-QBxXYfP4o8dgde1sB1jvo61KtR4Ofvoeerm9eb66Tx6f7h6uLh8TzbNpSLRgKeRMkzJl09zkUjMmpaxIlcqM5ISlXFLBaSxRmkyQyjCeUkGNphAF8T10ss5dOvs-gA9F13gNbat6sIMvoqFMEi5ZHtHTNaqd9d5BVSxd0yk3FpQUK-_FoizW3iN79BM7lB2YP_JXdASO14Adlv_kfAOzyYwf</recordid><startdate>20190201</startdate><enddate>20190201</enddate><creator>Jain, Rishu</creator><creator>Sharma, Deepak</creator><creator>Kumar, Rakesh</creator><creator>Kumar, Rajesh</creator><general>Oxford University Press</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>20190201</creationdate><title>Structural, kinetic and thermodynamic characterizations of SDS-induced molten globule state of a highly negatively charged cytochrome c</title><author>Jain, Rishu ; Sharma, Deepak ; Kumar, Rakesh ; Kumar, Rajesh</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c365t-c724e82c0b4258d89c22999f0f496080243917311737bd670fd234171dc1e0873</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2019</creationdate><topic>Cytochromes c - chemistry</topic><topic>Cytochromes c - metabolism</topic><topic>Kinetics</topic><topic>Protein Denaturation</topic><topic>Sodium Chloride - chemistry</topic><topic>Sodium Dodecyl Sulfate - chemistry</topic><topic>Thermodynamics</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Jain, Rishu</creatorcontrib><creatorcontrib>Sharma, Deepak</creatorcontrib><creatorcontrib>Kumar, Rakesh</creatorcontrib><creatorcontrib>Kumar, Rajesh</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of biochemistry (Tokyo)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Jain, Rishu</au><au>Sharma, Deepak</au><au>Kumar, Rakesh</au><au>Kumar, Rajesh</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Structural, kinetic and thermodynamic characterizations of SDS-induced molten globule state of a highly negatively charged cytochrome c</atitle><jtitle>Journal of biochemistry (Tokyo)</jtitle><addtitle>J Biochem</addtitle><date>2019-02-01</date><risdate>2019</risdate><volume>165</volume><issue>2</issue><spage>125</spage><epage>137</epage><pages>125-137</pages><issn>0021-924X</issn><eissn>1756-2651</eissn><abstract>Abstract
This study presents the structural, kinetic and thermodynamic characterizations of previously unknown submicellar concentrations of SDS-induced molten globule (MGSDS) state of a highly negatively charged base-denatured ferricytochrome c (UB-state) at pH ∼12.8 (±0.2). The far-UV CD, near-UV CD, ANS-fluorescence data of UB-state in the presence of different concentrations of SDS indicate that the submicellar concentrations of SDS (≤0.4 mM) transform the UB-state to MGSDS-state. The MGSDS-state has native-like α-helical secondary structure but lacks tertiary structure. The free energy change (ΔG°D) for UB→ MGSDS transition determined by far-UV CD (∼2.7 kcal mol−1) is slightly higher than those determined by fluorescence (∼2.0 kcal mol−1) at 25°C. At very low SDS and NaCl concentrations, the MGSDS-state undergoes cold denaturation. As SDS concentration is increased, the thermal denaturation temperature increases and the cold denaturation temperature decrease. Kinetic experiments involving the measurement of the CO-association rate to the base-denatured ferrocytochrome c at pH ≈12.8 (±0.2), 25°C indicate that the submicellar concentrations of SDS restrict the internal dynamics of base-denatured protein.</abstract><cop>England</cop><pub>Oxford University Press</pub><pmid>30371870</pmid><doi>10.1093/jb/mvy087</doi><tpages>13</tpages></addata></record> |
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subjects | Cytochromes c - chemistry Cytochromes c - metabolism Kinetics Protein Denaturation Sodium Chloride - chemistry Sodium Dodecyl Sulfate - chemistry Thermodynamics |
title | Structural, kinetic and thermodynamic characterizations of SDS-induced molten globule state of a highly negatively charged cytochrome c |
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