Start2Fold

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

Entry STF0013

Equine lysozyme

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Protein information

Name of the protein: Lysozyme C, milk isozyme
Organism: Equus caballus (Horse)
Number of residues: 129
Related UniProt entry:   P11376 (Fragment: 1 - 129)
Related PDB entry:   2EQL

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Experiment sets

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EARLY

Method: HDX-folding competition by NMR

Conditions: pH 7.5; 25.0 Celsius; Probes: 46 amides + 3 Trp indole hydrogen atoms

Related publication:
 PMID 10369782

Experiment details: "Equine lysozyme denatured in 6 M GdnHCl was diluted rapidly with deuterated buffer solution at pH 7.5 and 25 °C, in order to initiate simultaneous hydrogen exchange and protein refolding. The intrinsic time constants for amide hydrogen exchange under these conditions are of the order of 10-200 ms. Exchange was allowed to occur for one minute, and was then quenched by decreasing the pH to 4.5."

Protection threshold: strong protection within 3.5 ms

Sequence: KVFSKCELAHKLKAQEMDGFGGYSLANWVCMAEYESNFNTRAFNGKNANGSSDYGLFQLNNKWWCKDNKRSSSNACNIMCSKLLDENIDDDISCAKRVVRDPKGMSAWKAWVKHCKDKDLSEYLASCNL
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EARLY residues

8: L; 9: A; 11: K; 12: L; 13: K; 28: W; 29: V; 31: M; 111: W; 115: C;
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INTERMEDIATE

Method: HDX-folding competition by NMR

Conditions: pH 7.5; 25.0 Celsius; Probes: 46 amides + 3 Trp indole hydrogen atoms

Related publication:
 PMID 10369782

Experiment details: "Equine lysozyme denatured in 6 M GdnHCl was diluted rapidly with deuterated buffer solution at pH 7.5 and 25 °C, in order to initiate simultaneous hydrogen exchange and protein refolding. The intrinsic time constants for amide hydrogen exchange under these conditions are of the order of 10-200 ms. Exchange was allowed to occur for one minute, and was then quenched by decreasing the pH to 4.5."

Protection threshold: strong protection within 10 ms

Sequence: KVFSKCELAHKLKAQEMDGFGGYSLANWVCMAEYESNFNTRAFNGKNANGSSDYGLFQLNNKWWCKDNKRSSSNACNIMCSKLLDENIDDDISCAKRVVRDPKGMSAWKAWVKHCKDKDLSEYLASCNL
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INTERMEDIATE residues

15: Q; 23: Y; 27: N; 34: Y; 35: E;
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LATE

Method: HDX-folding competition by NMR

Conditions: pH 7.5; 25.0 Celsius; Probes: 46 amides + 3 Trp indole hydrogen atoms

Related publication:
 PMID 10369782

Experiment details: "Equine lysozyme denatured in 6 M GdnHCl was diluted rapidly with deuterated buffer solution at pH 7.5 and 25 °C, in order to initiate simultaneous hydrogen exchange and protein refolding. The intrinsic time constants for amide hydrogen exchange under these conditions are of the order of 10-200 ms. Exchange was allowed to occur for one minute, and was then quenched by decreasing the pH to 4.5."

Protection threshold: strong protection within 50 ms

Sequence: KVFSKCELAHKLKAQEMDGFGGYSLANWVCMAEYESNFNTRAFNGKNANGSSDYGLFQLNNKWWCKDNKRSSSNACNIMCSKLLDENIDDDISCAKRVVRDPKGMSAWKAWVKHCKDKDLSEYLASCNL
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LATE residues

4: S; 38: F; 61: N; 108: W;
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STRONG

Method: Native exchange NMR

Conditions: pH 4.5; 25.0 Celsius; Probes: 67

Related publication:
 PMID 9180380

Experiment details: "Samples for hydrogen exchange studies of the native holo state of equine lysozyme were adjusted to pH 4.5 in H2O and freeze-dried. The protein was then dissolved in D2O in the presence of 10 mM CaCl2, and the apparent pH adjusted to 4.5 using 2HCl and NaO2H. Samples were allowed to undergo exchange at 25 °C for time intervals ranging from a few minutes to 63 days. At the end of each exchange period, phase-sensitive COSY spectra were acquired."

Protection threshold: log(P) > 5

Sequence: KVFSKCELAHKLKAQEMDGFGGYSLANWVCMAEYESNFNTRAFNGKNANGSSDYGLFQLNNKWWCKDNKRSSSNACNIMCSKLLDENIDDDISCAKRVVRDPKGMSAWKAWVKHCKDKDLSEYLASCNL
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STRONG residues

9: A; 11: K; 12: L; 13: K; 27: N; 28: W; 31: M; 32: A; 38: F; 39: N; 61: N; 65: C; 93: S; 94: C; 95: A;
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MEDIUM

Method: Native exchange NMR

Conditions: pH 4.5; 25.0 Celsius; Probes: 67

Related publication:
 PMID 9180380

Experiment details: "Samples for hydrogen exchange studies of the native holo state of equine lysozyme were adjusted to pH 4.5 in H2O and freeze-dried. The protein was then dissolved in D2O in the presence of 10 mM CaCl2, and the apparent pH adjusted to 4.5 using 2HCl and NaO2H. Samples were allowed to undergo exchange at 25 °C for time intervals ranging from a few minutes to 63 days. At the end of each exchange period, phase-sensitive COSY spectra were acquired."

Protection threshold: 4 < log(P) < 5

Sequence: KVFSKCELAHKLKAQEMDGFGGYSLANWVCMAEYESNFNTRAFNGKNANGSSDYGLFQLNNKWWCKDNKRSSSNACNIMCSKLLDENIDDDISCAKRVVRDPKGMSAWKAWVKHCKDKDLSEYLASCNL
 CLICK TO DOWNLOAD SEQUENCE IN FASTA

MEDIUM residues

8: L; 10: H; 29: V; 33: E; 34: Y; 35: E; 36: S; 37: N; 40: T; 42: A; 54: Y; 57: F; 58: Q; 59: L; 62: K; 63: W; 66: K; 83: L; 96: K; 111: W; 112: V;
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