Start2Fold

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

Entry STF0007

Bovine alpha-lactalbumin (BLA)

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Protein information

Name of the protein: Alpha-lactalbumin
Organism: Bos taurus (Bovine)
Number of residues: 123
Related UniProt entry:   P00711 (Fragment: 20 - 142)
Related PDB entry:   1HFZ

Visualize the data

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Experiment sets

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EARLY

Method: HDX-folding competition by NMR

Conditions: pH 7.5; 10.0 Celsius; Probes: 36

Related publication:
 PMID 10329172

Experiment details: "A 200 mg/ml sample (14.1 mM) of apo-BLA (denatured and passed through a Pharmacia PD10 column) in 4.5 M GuHCl (H2O) and 15 mM EDTA was diluted 15-fold in 5 mM Tris-2HCl (D2O) at pH 7.5. After five seconds the pH was jumped to 5.5 and the Ca2‡ concentration to 20 mM by mixing with an equal volume of buffer containing 40 mM sodium acetate and 40 mM CaCl2."

Protection threshold: residues whose amide H atoms exchange slowly in the kinetic molten globule

Sequence: MEQLTKCEVFRELKDLKGYGGVSLPEWVCTTFHTSGYDTQAIVQNNDSTEYGLFQINNKIWCKDDQNPHSSNICNISCDKFLDDDLTDDIVCVKKILDKVGINYWLAHKALCSEKLDQWLCEKL
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EARLY residues

9: V; 11: R; 12: E; 13: L; 28: V; 43: V; 51: Y; 54: F; 55: Q; 56: I; 59: K; 61: W; 62: C; 63: K; 74: C; 76: I; 80: K; 81: F; 82: L; 85: D; 90: I; 91: V; 93: V; 94: K; 95: K; 96: I; 97: L; 98: D; 99: K; 100: V;
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STRONG

Method: Native exchange NMR

Conditions: pH 6.25; 20.0 Celsius; Probes: 49

Related publication:
 PMID 10329172

Experiment details: "Rates of solvent exchange of amide hydrogen atoms in the native state of BLA were measured by recording 2D 15N-1H HSQC spectra as a function of time following dissolution of the protein in D2O at pH 6.25."

Protection threshold: log(P) > 5

Sequence: MEQLTKCEVFRELKDLKGYGGVSLPEWVCTTFHTSGYDTQAIVQNNDSTEYGLFQINNKIWCKDDQNPHSSNICNISCDKFLDDDLTDDIVCVKKILDKVGINYWLAHKALCSEKLDQWLCEKL
 CLICK TO DOWNLOAD SEQUENCE IN FASTA

STRONG residues

49: T; 50: E; 53: L; 56: I; 59: K; 63: K; 64: D; 76: I; 78: C; 80: K; 81: F; 82: L; 83: D; 89: D; 90: I; 91: V; 93: V; 94: K; 95: K; 96: I; 97: L; 98: D; 99: K;
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WEAK

Method: Native exchange NMR

Conditions: pH 6.25; 20.0 Celsius; Probes: 49

Related publication:
 PMID 10329172

Experiment details: "Rates of solvent exchange of amide hydrogen atoms in the native state of BLA were measured by recording 2D 15N-1H HSQC spectra as a function of time following dissolution of the protein in D2O at pH 6.25."

Protection threshold: log(P) < 5

Sequence: MEQLTKCEVFRELKDLKGYGGVSLPEWVCTTFHTSGYDTQAIVQNNDSTEYGLFQINNKIWCKDDQNPHSSNICNISCDKFLDDDLTDDIVCVKKILDKVGINYWLAHKALCSEKLDQWLCEKL
 CLICK TO DOWNLOAD SEQUENCE IN FASTA

WEAK residues

9: V; 10: F; 11: R; 13: L; 27: W; 28: V; 41: A; 42: I; 43: V; 51: Y; 52: G; 54: F; 55: Q; 58: N; 61: W; 62: C; 73: I; 74: C; 77: S; 87: T; 88: D; 92: C; 100: V; 101: G; 102: I; 105: W;
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STRONG

Method: Native exchange in partially folded state by NMR

Conditions: pH 2.0; 20.0 Celsius; Probes: 36

Related publication:
 PMID 10329172

Experiment details: "BLA was dissolved in D2O at pH 2.0 for various lengths of time, the solvent exchange was quenched by freezing and the samples were lyophilized. The samples were subsequently analysed under native conditions by re-dissolving in native buffer at pH 5.5 in the presence of Ca2+‡."

Protection threshold: P > 4

Sequence: MEQLTKCEVFRELKDLKGYGGVSLPEWVCTTFHTSGYDTQAIVQNNDSTEYGLFQINNKIWCKDDQNPHSSNICNISCDKFLDDDLTDDIVCVKKILDKVGINYWLAHKALCSEKLDQWLCEKL
 CLICK TO DOWNLOAD SEQUENCE IN FASTA

STRONG residues

93: V; 94: K; 95: K; 96: I; 100: V; 101: G; 105: W;
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MEDIUM

Method: Native exchange in partially folded state by NMR

Conditions: pH 2.0; 20.0 Celsius; Probes: 36

Related publication:
 PMID 10329172

Experiment details: "BLA was dissolved in D2O at pH 2.0 for various lengths of time, the solvent exchange was quenched by freezing and the samples were lyophilized. The samples were subsequently analysed under native conditions by re-dissolving in native buffer at pH 5.5 in the presence of Ca2+‡."

Protection threshold: 2.5 < P < 4

Sequence: MEQLTKCEVFRELKDLKGYGGVSLPEWVCTTFHTSGYDTQAIVQNNDSTEYGLFQINNKIWCKDDQNPHSSNICNISCDKFLDDDLTDDIVCVKKILDKVGINYWLAHKALCSEKLDQWLCEKL
 CLICK TO DOWNLOAD SEQUENCE IN FASTA

MEDIUM residues

27: W; 62: C; 91: V; 92: C; 99: K;
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WEAK

Method: Native exchange in partially folded state by NMR

Conditions: pH 2.0; 20.0 Celsius; Probes: 36

Related publication:
 PMID 10329172

Experiment details: "BLA was dissolved in D2O at pH 2.0 for various lengths of time, the solvent exchange was quenched by freezing and the samples were lyophilized. The samples were subsequently analysed under native conditions by re-dissolving in native buffer at pH 5.5 in the presence of Ca2+‡."

Protection threshold: 0 < P < 2.5

Sequence: MEQLTKCEVFRELKDLKGYGGVSLPEWVCTTFHTSGYDTQAIVQNNDSTEYGLFQINNKIWCKDDQNPHSSNICNISCDKFLDDDLTDDIVCVKKILDKVGINYWLAHKALCSEKLDQWLCEKL
 CLICK TO DOWNLOAD SEQUENCE IN FASTA

WEAK residues

9: V; 10: F; 11: R; 13: L; 14: K; 28: V; 42: I; 43: V; 51: Y; 54: F; 56: I; 59: K; 61: W; 63: K; 73: I; 74: C; 76: I; 81: F; 82: L; 87: T; 90: I; 91: V; 97: L; 98: D; 102: I;
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STRONG

Method: Native exchange NMR

Conditions: pH 7.1; 20.0 Celsius; Probes: 24

Related publication:
 PMID 11273708

Experiment details: "A sample of authentic BLA from Sigma (concentration 1.4 mM) for hydrogen exchange was dissolved in 15 mM sodium phosphate buffer in H2O (pH 7.1) containing 20 mM EDTA, and 0.5 M NaCl. After lyophilisation, hydrogen exchange was initiated by dissolving the sample in 100 mM deuterated imidazole buffer (pH 7.1), containing a further 5 mM EDTA. Phase-sensitive COSY spectra 37 were collected on a home-built 500 MHz spectrometer, at 20 °C. A sweep width of 7518 was used in both dimensions. A series of seven COSY spectra was collected with 256 complex t1 increments of 1024 complex points; 32 scans were recorded per t1 increment. The acquisition time was 5.2 hours per spectrum. The data were zero-filled twice to 1 K in the t1 dimension, and zero-filled once to 2 K in the t2 dimension. Resolution enhancement was by trapezoidal and exponential multiplication in the t2 dimension and by trapezoidal multiplication in the t1 dimension."

Protection threshold: log(P) > 4

Sequence: MEQLTKCEVFRELKDLKGYGGVSLPEWVCTTFHTSGYDTQAIVQNNDSTEYGLFQINNKIWCKDDQNPHSSNICNISCDKFLDDDLTDDIVCVKKILDKVGINYWLAHKALCSEKLDQWLCEKL
 CLICK TO DOWNLOAD SEQUENCE IN FASTA

STRONG residues

13: L; 28: V; 38: D; 43: V; 54: F; 55: Q; 56: I; 58: N; 62: C; 63: K; 74: C; 76: I; 80: K; 81: F; 82: L; 89: D; 95: K; 97: L;
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