Start2Fold

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

Entry STF0005

Bovine beta-lactoglobulin (beta-LG)

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Protein information

Name of the protein: Beta-lactoglobulin
Organism: Bos taurus (Bovine)
Number of residues: 162
Related UniProt entry:   P02754 (Fragment: 17 - 178)
Related PDB entry:   3NPO

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Experiment sets

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EARLY

Method: Pulse labeling HDX NMR

Conditions: pH 8.3; 6.0 Celsius; Probes: 80

Related publication:
 PMID 10686102

Experiment details: "The H/2H exchange pulse labeling experiments were done manually at 6 C. The refolding of fully deuterated b-lactoglobulin was initiated from the TFE state in 40 % TFE (pH 3.0) or the Gdn-HCl-state in 6 MGdn-HCl (pH 3.0) by manually diluting 20-fold into D2O. At this stage, the pDr was confirmed to be 3.0. After the selected times (10, 30, 100 and 300 seconds) of refolding, the H/D exchange pulse labeling was carried out at pDr 8.3 by adding 1/100 volume of 250 mM Tris/D2O. For the refolding time zero at pDr 3.0, we directly diluted the denatured protein at pH 3.0 to the pulse labeling buffer at pDr 8.3. After an exchange pulse of 5 seconds at pDr 8.3, the reaction was quenched by lowering the pDr to 3.0. The denaturants and Tris were immediately removed by successive concentration/dilution of protein with 10 mM 2HCl. Throughout the procedures, the temperature of the solution was kept at 6 C. Then,HSQC spectra were measured to determine the extent of exchange at 25 C. The experiments were repeated twice and the results were similar within the error of +/-10%."

Protection threshold: proton occupancy > 0.7

Sequence: LIVTQTMKGLDIQKVAGTWYSLAMAASDISLLDAQSAPLRVYVEELKPTPEGDLEILLQKWENDECAQKKIIAEKTKIPAVFKIDALNENKVLVLDTDYKKYLLFCMENSAEPEQSLVCQCLVRTPEVDDEALEKFDKALKALPMHIRLSFNPTQLEEQCHI
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EARLY residues

37: A; 54: L; 56: I; 57: L; 58: L; 61: W; 65: E; 92: V; 94: V; 102: Y; 103: L; 104: L; 105: F; 106: C; 107: M; 117: L; 118: V; 119: C; 120: Q; 122: L; 123: V; 127: E; 139: A; 140: L; 143: L; 145: M; 147: I;
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EARLY

Method: Burst phase labeling HDX NMR

Conditions: pH 7.0-10.7; 20.0 Celsius; Probes: 80

Related publication:
 PMID 11175905

Experiment details: "Burst-phase labeling experiments at variable pulse pH and a constant refolding/exchange competition time of 1.8 ms"

Protection threshold: P > 10

Sequence: LIVTQTMKGLDIQKVAGTWYSLAMAASDISLLDAQSAPLRVYVEELKPTPEGDLEILLQKWENGECAQKKIIAEKTKIPAVFKIDALNENKVLVLDTDYKKYLLFCMENSAEPEQSLACQCLVRTPEVDDEALEKFDKALKALPMHIRLSFNPTQLEEQCHI
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EARLY residues

93: L; 100: K; 106: C; 107: M; 117: L; 122: L; 123: V; 132: A; 133: L; 139: A; 141: K;
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INTERMEDIATE

Method: Burst phase labeling HDX NMR

Conditions: pH 7.0-10.7; 20.0 Celsius; Probes: 80

Related publication:
 PMID 11175905

Experiment details: "Burst-phase labeling experiments at variable pulse pH and a constant refolding/exchange competition time of 1.8 ms"

Protection threshold: 5 < P < 10

Sequence: LIVTQTMKGLDIQKVAGTWYSLAMAASDISLLDAQSAPLRVYVEELKPTPEGDLEILLQKWENGECAQKKIIAEKTKIPAVFKIDALNENKVLVLDTDYKKYLLFCMENSAEPEQSLACQCLVRTPEVDDEALEKFDKALKALPMHIRLSFNPTQLEEQCHI
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INTERMEDIATE residues

12: I; 21: S; 91: K; 118: A; 119: C; 120: Q; 129: D; 130: D;
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LATE

Method: Burst phase labeling HDX NMR

Conditions: pH 7.0-10.7; 20.0 Celsius; Probes: 80

Related publication:
 PMID 11175905

Experiment details: "Burst-phase labeling experiments at variable pulse pH and a constant refolding/exchange competition time of 1.8 ms"

Protection threshold: 2 < P < 5

Sequence: LIVTQTMKGLDIQKVAGTWYSLAMAASDISLLDAQSAPLRVYVEELKPTPEGDLEILLQKWENGECAQKKIIAEKTKIPAVFKIDALNENKVLVLDTDYKKYLLFCMENSAEPEQSLACQCLVRTPEVDDEALEKFDKALKALPMHIRLSFNPTQLEEQCHI
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LATE residues

14: K; 17: G; 18: T; 19: W; 90: N; 109: N; 116: S;
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STRONG

Method: Native exchange NMR

Conditions: pH 2.5; 45.0 Celsius; Probes: every amide site

Related publication:
 PMID 10686102

Experiment details: "Corresponds to PDB code 3BLG! The H/D exchange was performed at 45 °C and pD 2.5 by dissolving the lyophilized protein into 10 mM DCl at a protein concentration of 10 mg/ml. The reaction was monitored by recording a series of 15N-1H HSQC spectra over four days."

Protection threshold: P > 10000

Sequence: LIVTQTMKGLDIQKVAGTWYSLAMAASDISLLDAQSAPLRVYVEELKPTPEGDLEILLQKWENDECAQKKIIAEKTKIPAVFKIDALNENKVLVLDTDYKKYLLFCMENSAEPEQSLVCQCLVRTPEVDDEALEKFDKALKALPMHIRLSFNPTQLEEQCHI
 CLICK TO DOWNLOAD SEQUENCE IN FASTA

STRONG residues

15: V; 20: Y; 24: M; 54: L; 56: I; 59: Q; 91: K; 92: V; 93: L; 94: V; 95: L; 103: L; 104: L; 105: F; 106: C; 107: M; 108: E; 115: Q; 118: V; 119: C; 120: Q; 122: L; 123: V; 136: F;
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MEDIUM

Method: Native exchange NMR

Conditions: pH 2.5; 45.0 Celsius; Probes: every amide site

Related publication:
 PMID 10686102

Experiment details: "Corresponds to PDB code 3BLG! The H/D exchange was performed at 45 °C and pD 2.5 by dissolving the lyophilized protein into 10 mM DCl at a protein concentration of 10 mg/ml. The reaction was monitored by recording a series of 15N-1H HSQC spectra over four days."

Protection threshold: 5000 < P < 10000

Sequence: LIVTQTMKGLDIQKVAGTWYSLAMAASDISLLDAQSAPLRVYVEELKPTPEGDLEILLQKWENDECAQKKIIAEKTKIPAVFKIDALNENKVLVLDTDYKKYLLFCMENSAEPEQSLVCQCLVRTPEVDDEALEKFDKALKALPMHIRLSFNPTQLEEQCHI
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MEDIUM residues

19: W; 23: A; 25: A; 43: V; 67: A; 84: I; 121: C;
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WEAK

Method: Native exchange NMR

Conditions: pH 2.5; 45.0 Celsius; Probes: every amide site

Related publication:
 PMID 10686102

Experiment details: "Corresponds to PDB code 3BLG! The H/D exchange was performed at 45 °C and pD 2.5 by dissolving the lyophilized protein into 10 mM DCl at a protein concentration of 10 mg/ml. The reaction was monitored by recording a series of 15N-1H HSQC spectra over four days."

Protection threshold: 1000 < P < 5000

Sequence: LIVTQTMKGLDIQKVAGTWYSLAMAASDISLLDAQSAPLRVYVEELKPTPEGDLEILLQKWENDECAQKKIIAEKTKIPAVFKIDALNENKVLVLDTDYKKYLLFCMENSAEPEQSLVCQCLVRTPEVDDEALEKFDKALKALPMHIRLSFNPTQLEEQCHI
 CLICK TO DOWNLOAD SEQUENCE IN FASTA

WEAK residues

16: A; 32: L; 41: V; 46: L; 57: L; 58: L; 71: I; 73: A; 74: E; 81: V; 82: F; 90: N; 102: Y; 109: N; 139: A; 140: L; 143: L;
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