Related ArticlesReductive cleavage and regeneration of the disulfide bonds in Streptomyces subtilisin inhibitor (SSI) as studied by the carbonyl 13C NMR resonances of cysteinyl residues.
J Biomol NMR. 1991 May;1(1):49-64
Authors: Uchida K, Miyake Y, Kainosho M
Four enhanced carbonyl carbon resonances were observed when Streptomyces subtilisin inhibitor (SSI) was labeled by incorporating specifically labeled [1-13C]Cys. The 13C signals were assigned by the 15N, 13C double-labeling method along with site-specific mutagenesis. Changes in the spectrum of the labeled protein ([C]SSI) were induced by reducing the disulfide bonds with various amounts of dithiothreitol (DTT). The results indicate that, in the absence of denaturant, the Cys71-Cys101 disulfide bond of each SSI subunit can be reduced selectively. This disulfide bond, which is in the vicinity of the reactive site scissile bond Met73-Val74, is more accessible to solvent than the other disulfide bond, Cys35-Cys50, which is embedded in the interior of SSI. This half-reduced SSI had 65% of the inhibitory activity of native SSI and maintained a conformation similar to that of the fully oxidized SSI. Reoxidation of the half reduced-folded SSI by air regenerates fully active SSI which is indistinguishable with intact SSI by NMR. In the presence of 3 M guanidine hydrochloride (GuHCl), however, both disulfide bonds of each SSI subunit were readily reduced by DTT. The fully reduced-unfolded SSI spontaneously refolded into a native-like structure (fully reduced-folded state), as evidenced by the Cys carbonyl carbon chemical shifts, upon removing GuHCl and DTT from the reaction mixture. The time course of disulfide bond regeneration from this state by air oxidation was monitored by following the NMR spectral changes and the results indicated that the disulfide bond between Cys71 and Cys101 regenerates at a much faster rate than that between Cys35 and Cys50.
Conformational analysis by quantitative NOE measurements of the β-proton pairs across individual disulfide bonds in proteins
Conformational analysis by quantitative NOE measurements of the β-proton pairs across individual disulfide bonds in proteins
Abstract NOEs between the β-protons of cysteine residues across disulfide bonds in proteins provide direct information on the connectivities and conformations of these important cross-links, which are otherwise difficult to investigate. With conventional -proteins, however, fast spin diffusion processes mediated by strong dipolar interactions between geminal β-protons prohibit the quantitative measurements and thus the analyses of long-range NOEs across...
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[NMR paper] Disulfide bonds in homo- and heterodimers of EF-hand subdomains of calbindin D9k: sta
Disulfide bonds in homo- and heterodimers of EF-hand subdomains of calbindin D9k: stability, calcium binding, and NMR studies.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--www3.interscience.wiley.com-aboutus-images-wiley_interscience_pubmed_logo_FREE_120x27.gif http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--www.pubmedcentral.nih.gov-corehtml-pmc-pmcgifs-pubmed-pmc.gif Related Articles Disulfide bonds in homo- and heterodimers of EF-hand subdomains of calbindin D9k: stability, calcium binding, and NMR studies.
Protein Sci. 1993 Jun;2(6):985-1000
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[NMR paper] NMR and isotopic exchange studies of the site of bond cleavage in the MutT reaction.
NMR and isotopic exchange studies of the site of bond cleavage in the MutT reaction.
Related Articles NMR and isotopic exchange studies of the site of bond cleavage in the MutT reaction.
J Biol Chem. 1992 Aug 25;267(24):16939-42
Authors: Weber DJ, Bhatnagar SK, Bullions LC, Bessman MJ, Mildvan AS
The MutT protein, which prevents AT----CG transversions during DNA replication, hydrolyzes nucleoside triphosphates to yield nucleoside monophosphates and pyrophosphate. The hydrolysis of dGTP by the MutT protein in H(2)18O-enriched water, when...
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[NMR paper] H-NMR studies of native and fragmented subtilisin Carlsberg.
H-NMR studies of native and fragmented subtilisin Carlsberg.
Related Articles H-NMR studies of native and fragmented subtilisin Carlsberg.
Biochim Biophys Acta. 1992 Feb 13;1119(1):39-44
Authors: Consonni R, Molinari H, Greco F, Zannoni G, Zetta L, Carrea G, Riva S
NMR studies have been carried out on subtilisin Carlsberg in order to identify the sharp resonances observed in the proton spectra of the enzyme dissolved in aqueous solution. NMR spectra, obtained with the combination of spin-echo and selective excitation sequences, from both the...
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[NMR paper] Characterization by NMR of the heme-myoglobin adduct formed during the reductive meta
Characterization by NMR of the heme-myoglobin adduct formed during the reductive metabolism of BrCCl3. Covalent bonding of the proximal histidine to the ring I vinyl group.
Related Articles Characterization by NMR of the heme-myoglobin adduct formed during the reductive metabolism of BrCCl3. Covalent bonding of the proximal histidine to the ring I vinyl group.
J Biol Chem. 1991 Feb 15;266(5):3208-14
Authors: Osawa Y, Highet RJ, Bax A, Pohl LR
The reductive debromination of BrCCl3 by ferrous deoxymyoglobin leads to the covalent bonding of the...
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[NMR paper] Cold denaturation and heat denaturation of Streptomyces subtilisin inhibitor. 2. 1H N
Cold denaturation and heat denaturation of Streptomyces subtilisin inhibitor. 2. 1H NMR studies.
Related Articles Cold denaturation and heat denaturation of Streptomyces subtilisin inhibitor. 2. 1H NMR studies.
Biochemistry. 1991 Nov 26;30(47):11313-20
Authors: Tamura A, Kimura K, Akasaka K
Structural transitions of the protein Streptomyces subtilisin inhibitor (SSI) from the native state to the cold-denatured and heat-denatured states were studied by 1H NMR spectroscopy in the temperature range from -10 to 60 degrees C in the acidic pH range....
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[NMR paper] Cold denaturation and heat denaturation of Streptomyces subtilisin inhibitor. 2. 1H N
Cold denaturation and heat denaturation of Streptomyces subtilisin inhibitor. 2. 1H NMR studies.
Related Articles Cold denaturation and heat denaturation of Streptomyces subtilisin inhibitor. 2. 1H NMR studies.
Biochemistry. 1991 Nov 26;30(47):11313-20
Authors: Tamura A, Kimura K, Akasaka K
Structural transitions of the protein Streptomyces subtilisin inhibitor (SSI) from the native state to the cold-denatured and heat-denatured states were studied by 1H NMR spectroscopy in the temperature range from -10 to 60 degrees C in the acidic pH range....
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[NMR paper] 13C-NMR of Clostridium pasteurianum ferredoxin after reductive methylation of the ami
13C-NMR of Clostridium pasteurianum ferredoxin after reductive methylation of the amines using formaldehyde.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--linkinghub.elsevier.com-ihub-images-PubMedLink.gif Related Articles 13C-NMR of Clostridium pasteurianum ferredoxin after reductive methylation of the amines using formaldehyde.
Biochim Biophys Acta. 1990 Apr 19;1038(2):146-51
Authors: Gluck M, Sweeney WV
Clostridium pasteurianum 2(4Fe-4S) ferredoxin has been reductively methylated using formaldehyde and sodium cyanoborohydride....