ジャーナル: J Biol Chem / 年: 2015 タイトル: Olfactomedin-1 Has a V-shaped Disulfide-linked Tetrameric Structure. 著者: Matti F Pronker / Trusanne G A A Bos / Thomas H Sharp / Dominique M E Thies-Weesie / Bert J C Janssen / 要旨: Olfactomedin-1 (Olfm1; also known as noelin and pancortin) is a member of the olfactomedin domain-containing superfamily and a highly expressed neuronal glycoprotein important for nervous system ...Olfactomedin-1 (Olfm1; also known as noelin and pancortin) is a member of the olfactomedin domain-containing superfamily and a highly expressed neuronal glycoprotein important for nervous system development. It binds a number of secreted proteins and cell surface-bound receptors to induce cell signaling processes. Using a combined approach of x-ray crystallography, solution scattering, analytical ultracentrifugation, and electron microscopy we determined that full-length Olfm1 forms disulfide-linked tetramers with a distinctive V-shaped architecture. The base of the "V" is formed by two disulfide-linked dimeric N-terminal domains. Each of the two V legs consists of a parallel dimeric disulfide-linked coiled coil with a C-terminal β-propeller dimer at the tips. This agrees with our crystal structure of a C-terminal coiled-coil segment and β-propeller combination (Olfm1(coil-Olf)) that reveals a disulfide-linked dimeric arrangement with the β-propeller top faces in an outward exposed orientation. Similar to its family member myocilin, Olfm1 is stabilized by calcium. The dimer-of-dimers architecture suggests a role for Olfm1 in clustering receptors to regulate signaling and sheds light on the conformation of several other olfactomedin domain family members.
CORRESPONDS TO UNIPROT ENTRY O88998, BUT WITH MUTATION A329T
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実験情報
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実験
実験
手法: X線回折 / 使用した結晶の数: 1
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試料調製
結晶
マシュー密度: 2.9 Å3/Da / 溶媒含有率: 57 % / 解説: NONE
結晶化
温度: 293 K / 手法: 蒸気拡散法, シッティングドロップ法 / pH: 8.5 詳細: SITTING DROP AT 293 K, MIXING PROTEIN AT 6 MG/ML 1:1 WITH PRECIPITANT SOLUTION: 1M LICL, 20% PEG6000 (W/V) AND 100 MM TRIS PH8.5
解像度: 2.4→50.318 Å / SU ML: 0.33 / σ(F): 1.92 / 位相誤差: 32.14 / 立体化学のターゲット値: ML 詳細: RESIDUES 17-210, 339-352 AND 478-487 NOT MODELED BECAUSE OF DISORDER OR LIMITED PROTEOLYSIS
Rfactor
反射数
%反射
Rfree
0.2578
2420
5 %
Rwork
0.237
-
-
obs
0.238
26030
94.72 %
溶媒の処理
減衰半径: 0.9 Å / VDWプローブ半径: 1.11 Å / 溶媒モデル: FLAT BULK SOLVENT MODEL / Bsol: 0.4242 Å2 / ksol: 1.3585 e/Å3
原子変位パラメータ
Biso mean: 69.76 Å2
精密化ステップ
サイクル: LAST / 解像度: 2.4→50.318 Å
タンパク質
核酸
リガンド
溶媒
全体
原子数
4119
0
159
26
4304
拘束条件
Refine-ID
タイプ
Dev ideal
数
X-RAY DIFFRACTION
f_bond_d
0.002
4396
X-RAY DIFFRACTION
f_angle_d
0.555
5980
X-RAY DIFFRACTION
f_dihedral_angle_d
11.226
1555
X-RAY DIFFRACTION
f_chiral_restr
0.025
664
X-RAY DIFFRACTION
f_plane_restr
0.001
750
LS精密化 シェル
解像度 (Å)
Rfactor Rfree
Num. reflection Rfree
Rfactor Rwork
Num. reflection Rwork
Refine-ID
% reflection obs (%)
2.4-2.449
0.3722
134
0.3624
2730
X-RAY DIFFRACTION
95
2.449-2.5023
0.4303
146
0.3436
2690
X-RAY DIFFRACTION
96
2.5023-2.5605
0.3318
142
0.3369
2646
X-RAY DIFFRACTION
94
2.5605-2.6245
0.3811
135
0.3323
2690
X-RAY DIFFRACTION
95
2.6245-2.6955
0.352
130
0.3324
2535
X-RAY DIFFRACTION
90
2.6955-2.7748
0.3225
141
0.3161
2725
X-RAY DIFFRACTION
96
2.7748-2.8643
0.3442
142
0.3037
2689
X-RAY DIFFRACTION
96
2.8643-2.9667
0.3378
131
0.2996
2785
X-RAY DIFFRACTION
96
2.9667-3.0854
0.2899
145
0.2943
2667
X-RAY DIFFRACTION
95
3.0854-3.2258
0.3197
117
0.2662
2639
X-RAY DIFFRACTION
94
3.2258-3.3959
0.2537
141
0.2538
2639
X-RAY DIFFRACTION
93
3.3959-3.6086
0.2866
165
0.2417
2684
X-RAY DIFFRACTION
96
3.6086-3.8871
0.2296
174
0.2288
2673
X-RAY DIFFRACTION
96
3.8871-4.2781
0.196
136
0.1985
2686
X-RAY DIFFRACTION
95
4.2781-4.8967
0.2117
142
0.1673
2747
X-RAY DIFFRACTION
95
4.8967-6.1676
0.1988
148
0.2002
2687
X-RAY DIFFRACTION
96
6.1676-50.3286
0.2411
151
0.2063
2657
X-RAY DIFFRACTION
95
精密化 TLS
手法: refined / Refine-ID: X-RAY DIFFRACTION
ID
L11 (°2)
L12 (°2)
L13 (°2)
L22 (°2)
L23 (°2)
L33 (°2)
S11 (Å °)
S12 (Å °)
S13 (Å °)
S21 (Å °)
S22 (Å °)
S23 (Å °)
S31 (Å °)
S32 (Å °)
S33 (Å °)
T11 (Å2)
T12 (Å2)
T13 (Å2)
T22 (Å2)
T23 (Å2)
T33 (Å2)
Origin x (Å)
Origin y (Å)
Origin z (Å)
1
5.8329
4.093
6.7257
9.9925
5.6294
7.9202
0.4786
-2.2111
0.2765
1.8326
-0.7606
0.847
0.8701
-2.9152
0.2663
0.6939
-0.1441
0.0485
1.2672
-0.0526
0.7499
86.1233
10.399
30.3389
2
5.9036
0.9248
0.4192
2.7137
0.3081
4.5075
0.205
0.2551
-0.233
0.0529
-0.0827
-0.1353
-0.0679
0.1215
-0.1078
0.2917
-0.057
-0.0231
0.4722
-0.0098
0.4499
64.9776
8.611
24.9528
3
9.5538
-6.9844
6.7089
2.1308
-8.4107
2.0268
-0.6208
-0.64
0.7762
1.3958
-0.509
-0.9618
-1.3306
1.8112
1.025
0.7603
-0.2699
-0.1281
1.2714
-0.0309
0.7559
95.3386
12.1812
29.6563
4
5.7316
0.6697
2.9256
3.0028
0.274
4.436
0.153
-0.1596
-0.1011
0.1024
-0.1614
-0.0524
-0.0019
-0.1596
0.0007
0.2926
-0.1027
0.0244
0.5492
-0.0484
0.4439
112.2599
15.3075
15.2269
精密化 TLSグループ
ID
Refine-ID
Refine TLS-ID
Selection details
1
X-RAY DIFFRACTION
1
CHAIN 'A' AND (RESID211THROUGH227 )
2
X-RAY DIFFRACTION
2
CHAIN 'A' AND (RESID228THROUGH477 ) ORCHAIN 'S' AND (RESID44)
3
X-RAY DIFFRACTION
3
CHAIN 'B' AND (RESID210THROUGH227 )
4
X-RAY DIFFRACTION
4
CHAIN 'B' AND (RESID228THROUGH480 ) ORCHAIN 'S' AND (RESID45)