The structures are that of the fully oleate-liganded LFABP (protein-ligand ratio 1:2). However, ...The structures are that of the fully oleate-liganded LFABP (protein-ligand ratio 1:2). However, they are drived from information pertaining protein alone. A major form of the complex coordinates is deposited separately.
Fattyacid-bindingprotein, liver / L-FABP / Z-protein / Squalene- and sterol-carrier protein / SCP / p14
Mass: 14293.544 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Rattus norvegicus (Norway rat) / Gene: Fabp1 / Production host: Escherichia coli (E. coli) / Strain (production host): BL21(DE3) / Variant (production host): pLys / References: UniProt: P02692
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Experimental details
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Experiment
Experiment
Method: SOLUTION NMR Details: The structures are that of the fully oleate-liganded LFABP (protein-ligand ratio 1:2). However, they are drived from information pertaining protein alone. A major form of the complex ...Details: The structures are that of the fully oleate-liganded LFABP (protein-ligand ratio 1:2). However, they are drived from information pertaining protein alone. A major form of the complex coordinates is deposited separately.
NMR experiment
Conditions-ID
Experiment-ID
Solution-ID
Type
1
1
3
2D 1H-15N HSQC
1
2
1
3DCBCA(CO)NH
1
3
1
3D HN(CA)CB
2
4
2
3D 1H-13C NOESY
2
5
2
4D 1H-13C NOESY
1
6
1
3D 1H-15N NOESY
1
7
1
4D 1H-13C-15N NOESY
1
8
2
3D (H)CCH-TOCSY
1
9
3
3D HNHA
1
10
1
3DC(CO)NH
1
11
1
3DH(CO)NH
1
12
1
3D HNCO
1
13
1
3DCBCACOHA
1
14
1
2D (HB)CB(CD)HD
1
15
1
2D (HB)CB(CDCE)HE
1
16
1
2DCG(CB)HB
1
17
1
2DCG(CD)HD
1
18
1
2DCG(CDCE)HE
1
19
1
3D CT-HMQC-COSY
1
20
1
2DhisHCN
1
21
1
2D 1H-15N HSQC
NMR details
Text: RDC data from sample 4 are not used (and hence not reported) because the medium (C12E5) competes with LFABP for oleate
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Sample preparation
Details
Solution-ID
Contents
Solvent system
1
0.7-1.0 mM [U-99% 13C; U-99% 15N] LFABP, 50 mM sodium phosphate, 0.02 % sodium azide, 5 uM EDTA, 5 % D2O, 3 eq oleate, 100 mM sodium chloride, 95% H2O/5% D2O
95% H2O/5% D2O
2
0.7-1.0 mM [U-99% 13C; U-99% 15N] LFABP, 50 mM sodium phosphate, 0.02 % sodium azide, 5 uM EDTA, 98 % D2O, 3 eq oleate, 100 mM sodium chloride, 100% D2O
100% D2O
3
0.5-1.3 mM [U-99% 15N] LFABP, 50 mM sodium phosphate, 0.02 % sodium azide, 5 uM EDTA, 5 % D2O, 3 eq [U-99% 13C] oleate, 100 mM sodium chloride, 95% H2O/5% D2O
95% H2O/5% D2O
4
0.3 mM [U-99% 15N] LFABP, 50 mM sodium phosphate, 0.02 % sodium azide, 5 uM EDTA, 5 % D2O, 5 % C12E5, 3 eq [U-99% 13C] oleate, 100 mM sodium chloride, 95% H2O/5% D2O