Start2Fold

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

Entry STF0014

Human lysozyme

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Protein information

Name of the protein: Lysozyme C
Organism: Homo sapiens
Number of residues: 130
Related UniProt entry:   P61626 (Fragment: 19 - 148)
Related PDB entry:   1LZ1

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Experiment sets

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EARLY

Method: Pulse labeling HDX NMR

Conditions: pH 5.3; 20.0 Celsius; Probes: 47

Related publication:
 PMID 8180215

Experiment details: "A QFM5 module was used to dilute a solution of human lysozyme in 6 M GuDCl /D2O (1 vol) with 20 mM sodium acetate in H2O, pH 5.5 (10 vol), to initiate the refolding reaction. The intrinsic rate of amide hydrogen exchange at 20 °C at the final refolding pH of 5.3 is approximately 10 s, and thus no significant exchange of amide deuterons occurs over the time course of folding (0-500 ms) monitored in this phase. At different times following the initiation of refolding, the pH of the refolding medium was jumped to 9.3 by dilution with 200 mM sodium borate buffer at pH 10.0 (5 volumes); this labeling pulse was applied for 8 ms."

Protection threshold: protection within 3.5 ms

Sequence: KVFERCELARTLKRLGMDGYRGISLANWMCLAKWESGYNTRATNYNAGDRSTDYGIFQINSRYWCNDGKTPGAVNACHLSCSALLQDNIADAVACAKRVVRDPQGIRAWVAWRNRCQNRDVRQYVQGCGV
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EARLY residues

8: L; 10: R; 11: T; 12: L; 13: K; 14: R; 29: M; 30: C; 31: L; 33: K; 34: W; 124: Y; 125: V;
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INTERMEDIATE

Method: Pulse labeling HDX NMR

Conditions: pH 5.3; 20.0 Celsius; Probes: 47

Related publication:
 PMID 8180215

Experiment details: "A QFM5 module was used to dilute a solution of human lysozyme in 6 M GuDCl /D2O (1 vol) with 20 mM sodium acetate in H2O, pH 5.5 (10 vol), to initiate the refolding reaction. The intrinsic rate of amide hydrogen exchange at 20 °C at the final refolding pH of 5.3 is approximately 10 s, and thus no significant exchange of amide deuterons occurs over the time course of folding (0-500 ms) monitored in this phase. At different times following the initiation of refolding, the pH of the refolding medium was jumped to 9.3 by dilution with 200 mM sodium borate buffer at pH 10.0 (5 volumes); this labeling pulse was applied for 8 ms."

Protection threshold: protection within 70 ms

Sequence: KVFERCELARTLKRLGMDGYRGISLANWMCLAKWESGYNTRATNYNAGDRSTDYGIFQINSRYWCNDGKTPGAVNACHLSCSALLQDNIADAVACAKRVVRDPQGIRAWVAWRNRCQNRDVRQYVQGCGV
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INTERMEDIATE residues

64: W; 65: C; 79: L; 91: D; 92: A; 93: V; 94: A; 95: C; 96: A; 97: K; 98: R; 100: V; 110: V; 112: W; 116: C;
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LATE

Method: Pulse labeling HDX NMR

Conditions: pH 5.3; 20.0 Celsius; Probes: 47

Related publication:
 PMID 8180215

Experiment details: "A QFM5 module was used to dilute a solution of human lysozyme in 6 M GuDCl /D2O (1 vol) with 20 mM sodium acetate in H2O, pH 5.5 (10 vol), to initiate the refolding reaction. The intrinsic rate of amide hydrogen exchange at 20 °C at the final refolding pH of 5.3 is approximately 10 s, and thus no significant exchange of amide deuterons occurs over the time course of folding (0-500 ms) monitored in this phase. At different times following the initiation of refolding, the pH of the refolding medium was jumped to 9.3 by dilution with 200 mM sodium borate buffer at pH 10.0 (5 volumes); this labeling pulse was applied for 8 ms."

Protection threshold: protection around 100 ms

Sequence: KVFERCELARTLKRLGMDGYRGISLANWMCLAKWESGYNTRATNYNAGDRSTDYGIFQINSRYWCNDGKTPGAVNACHLSCSALLQDNIADAVACAKRVVRDPQGIRAWVAWRNRCQNRDVRQYVQGCGV
 CLICK TO DOWNLOAD SEQUENCE IN FASTA

LATE residues

3: F; 38: Y; 39: N; 40: T; 41: R; 42: A; 44: N; 51: S; 53: D; 54: Y; 58: Q; 59: I; 60: N; 62: R; 66: N; 77: C; 84: L;
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