Start2Fold

The database of hydrogen/deuterium exchange data on protein folding and stability

Entry STF0020

SNase H124L

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Protein information

Name of the protein: Thermonuclease
Organism: Staphylococcus aureus
Number of residues: 149
Related UniProt entry:   P00644 (Fragment: 83 - 231)
Related PDB entry:   1JOO

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Experiment sets

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EARLY

Method: Pulse labeling HDX NMR

Conditions: pH 5.3; 15.0 Celsius; Probes: 60

Related publication:
 PMID 11742125

Experiment details: "Refolding of deuterated H124L SNase was initiated at 15°C by a 1:2 dilution of the unfolded protein with a refolding buffer containing 150 mM KCl and 75 mM sodium acetate (pH 6.8) such that the final refolding mixture had a pH* of 5.3 and contained 100 mM KCl and 50 mM sodium acetate. Refolding was allowed to continue for a variable length of time."

Protection threshold: P > 5

Sequence: ATSTKKLHKEPATLIKAIDGDTVKLMYKGQPMTFRLLLVDTPETKHPKKGVEKYGPEASAFTKKMVENAKKIEVEFDKGQRTDKYGRGLAYIYADGKMVNEALVRQGLAKVAYVYKPNNTHEQLLRKSEAQAKKEKLNIWSEDNADSGQ
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EARLY residues

23: V; 24: K; 25: L; 26: M; 27: Y; 32: M; 34: F; 35: R; 137: L;
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LATE

Method: Pulse labeling HDX NMR

Conditions: pH 5.3; 15.0 Celsius; Probes: 60

Related publication:
 PMID 11742125

Experiment details: "Refolding of deuterated H124L SNase was initiated at 15°C by a 1:2 dilution of the unfolded protein with a refolding buffer containing 150 mM KCl and 75 mM sodium acetate (pH 6.8) such that the final refolding mixture had a pH* of 5.3 and contained 100 mM KCl and 50 mM sodium acetate. Refolding was allowed to continue for a variable length of time."

Protection threshold: 2.5 > P < 5

Sequence: ATSTKKLHKEPATLIKAIDGDTVKLMYKGQPMTFRLLLVDTPETKHPKKGVEKYGPEASAFTKKMVENAKKIEVEFDKGQRTDKYGRGLAYIYADGKMVNEALVRQGLAKVAYVYKPNNTHEQLLRKSEAQAKKEKLNIWSEDNADSGQ
 CLICK TO DOWNLOAD SEQUENCE IN FASTA

LATE residues

37: L; 88: G; 91: Y; 97: K; 138: N; 139: I; 141: S;
 CLICK TO DOWNLOAD LIST OF RESIDUES

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STRONG

Method: Native exchange NMR

Conditions: pH 5.5; 37.0 Celsius; Probes: 100-105

Related publication:
 PMID 8218167

Experiment details: "Samples for exchange studies were prepared by dissolving the lyophilized protein in an aqueous solution (natural isotopic abundance) containing 50 mM succinate-d4 to a final concentration of 3.0 mM. Samples of the nuclease H124L-Ca2+-pdTp ternary complex contained an additional 9.0 mM pdTp and 18 mM CaCl2. The pH was adjusted to 5.1, and the samples were lyophilized to dryness. Immediately prior to NMR data acquisition, the samples were redissolved in 100% D2O and placed into the Bruker AM500 NMR spectrometer."

Protection threshold: G(op)(kcal/mol)=~G(global unfolding)

Sequence: ATSTKKLHKEPATLIKAIDGDTVKLMYKGQPMTFRLLLVDTPETKHPKKGVEKYGPEASAFTKKMVENAKKIEVEFDKGQRTDKYGRGLAYIYADGKMVNEALVRQGLAKVAYVYKPNNTHEQLLRKSEAQAKKEKLNIWSEDNADSGQ
 CLICK TO DOWNLOAD SEQUENCE IN FASTA

STRONG residues

22: T; 24: K; 25: L; 26: M; 34: F; 37: L;
 CLICK TO DOWNLOAD LIST OF RESIDUES