鍖栧浜よ伅(li谩n)绲�(ji茅)鍚堣唱(zh矛)璀滄妧琛�(sh霉)锛坈hemical cross-linking coupled with mass spectrometry锛孋XMS锛夋妧琛�(sh霉)鍦ㄨ繎鍗佸勾鍏�(n猫i)寰楀埌浜嗚摤鍕冪櫦(f膩)灞�銆傝寤ｆ硾鎳夌敤浜庤泲鐧借唱(zh矛)绲�(ji茅)妲�(g貌u)閼掑畾鍜岃泲鐧借唱(zh矛)鐩镐簰浣滅敤鍒嗘瀽銆傜洰鍓岰XMS鎶�琛�(sh霉)鍦ㄥ闅涙噳鐢ㄥ瓨鍦ㄤ竴瀹氱殑灞�闄愭�э細1銆佸京闆滄ǎ鍝佽檿鐞嗘晥鐜囧亸浣�锛�2銆佽兘澶犳彁渚涚殑浜よ伅(li谩n)淇℃伅鏈夐檺銆傝┎鎶�琛�(sh霉)鐧�(f膩)灞曠殑鏈�澶х摱闋稿凡缍�(j墨ng)楂旂従(xi脿n)鍦ㄦ湁闄愮殑鍖栧浜よ伅(li谩n)鍔戦伕鎿囧拰浜よ伅(li谩n)鏁堢巼涓�銆傜櫨闈堝▉浣滅偤灏堟キ(y猫)渚涙噳鍟嗭紝鎻愪緵鏂板瀷鍖栧浜よ伅(li谩n)鍔慙eiker鍜宒6-Leiker锛屽叿鏈夊彲瀵岄泦銆佸彲鍒囨柗銆佸彲瀹氶噺鐨勪綔鐢�[1]銆�
鈻� Leiker鍏锋湁楂樻晥鐨勫瘜闆嗗姛鑳斤紝姣斿偝绲�(t菕ng)浜よ伅(li谩n)鍔戞湁鏇撮珮鐨勬晥鐜�
Figure 1. Leiker(2138035) allowed near 100% enrichment of target peptides from a cross-linked ten-protein mixture diluted with increasing amounts of non-cross-linked E. coli lysates[1].
鈻� Leiker鍙互椤憲鎻愰珮鎵�鑳介憭瀹氬埌鐨勪氦鑱�(li谩n)鑲芥鏁�(sh霉)閲�
Figure 2. CXMS analyses of E. coli and C. elegans lysates. The best protein-protein interaction cluster extracted from the Leiker-identified or BS3(146482)-identified (Yang et al., 2012) [2]inter-links from E. coli whole-cell lysates[1].
鈻� Leiker鍙互閫茶瀹氶噺CXMS鐮旂┒
Figure 3. Quantitative CXMS analysis of the L7Ae-RNA complex. Reciprocal labeling of RNA-free (F) and RNA-bound (B) L7Ae with [d0]/[d6]-Leiker(2322681)[1].
References
[1] Tan D, Li Q, Zhang M J, et al. Trifunctional cross-linker for mapping protein-protein interaction networks and comparing protein conformational states[J]. Elife Sciences, 2016, 5.
[2] Yang B, Wu Y J, Zhu M, et al. Identification of cross-linked peptides from complex samples.[J]. Nature Methods, 2012, 9(9):904-906.
鈻� 鏂板瀷铔嬬櫧璩�(zh矛)浜よ伅(li谩n)鍔�
|
鐕熼姺绶ㄨ櫉
|
鐢�(ch菐n)鍝佸悕绋�
|
涓枃鍚嶇ū
|
鍖呰瑕�(gu墨)鏍�
|
|
2138035
|
Leiker, 95%
|
璩存皑閰哥壒鐣版�у彲瀵岄泦鍨嬩氦鑱�(li谩n)鍔�
|
5 mg
10 mg
|
|
2322681
|
d6-Leiker, 95%
|
d6-璩存皑閰哥壒鐣版�у彲瀵岄泦鍨嬩氦鑱�(li谩n)鍔�
|
5 mg
10 mg
|
鈻� 鍏朵粬浜よ伅(li谩n)鍔戠郴鍒楃敘(ch菐n)鍝�
|
鐢�(ch菐n)鍝佺法铏�
|
鐢�(ch菐n)鍝佸悕绋�
|
CAS
|
鍖呰瑕�(gu墨)鏍�
|
|
350636
|
4-Benzoylbenzoic acid N-succinimidyl ester, 97%
|
91990-88-4
|
50 mg
250 mg
|
|
146482
|
Bis(sulfosuccinimidyl) suberate sodium salt, BS3, 90%
|
82436-77-9
|
25 mg
100 mg
|
|
296329
|
Di(N-succinimidyl) 3,3'-dithiodipropionate, DTSP, 97%
|
57757-57-0
|
1 g
5 g
|
|
456454
|
Di(N-succinimidyl) glutarate, DSG, 97%
|
79642-50-5
|
50 mg
250 mg
|
|
253405
|
Di(N-succinimidyl) suberate, DSS, 98%
|
68528-80-3
|
1 g
5 g
|
|
1410146
|
Disuccinimidyl tartrate, DST, 95%
|
N/A
|
5 mg
25 mg
|
|
792543
|
3,3'-dithiobispropionic acid bis-sulfosuccinimidyl ester, DTSSP, 90%
|
81069-02-5
|
25 mg
100 mg
|
|
444282
|
N-Succinimidyl iodoacetate, 95%
|
39028-27-8
|
100 mg
500 mg
|
|
551313
|
Succinimidyl 3-(2-Pyridyldithio)Propionate, SPDP, 97%
|
68181-17-9
|
100 mg
|
鈻� 鐩搁棞(gu膩n)鐢�(ch菐n)鍝�
|
鐢�(ch菐n)鍝佺法铏�
|
鑻辨枃鍚嶇ū
|
涓枃鍚嶇ū
|
CAS
|
|
649777
|
Acrylamide/?Bis-?acrylamide Solution, 29:1, 40%
|
涓欑儻閰拌兒/闆欎笝鐑叞鑳烘憾娑�
|
N/A
|
|
398539
|
Aprotinin, 6000 U/mg
|
鎶戣偨閰�
|
9087-70-1
|
|
3922801
|
BCA Protein Assay Macro Kit
|
铔嬬櫧璩�(zh矛)瀹氶噺瑭﹀姂鐩�
|
N/A
|
|
266630
|
Benzamidine hydrochloride, 99%
|
鑻敳鑴掗菇閰搁菇
|
1670-14-0
|
|
1914018
|
CAPS Buffer
|
CAPS 绶╂矕娑�
|
N/A
|
|
327499
|
Coomassie Brilliant Blue G
|
鑰冮Μ鏂寒钘�G
|
6104-58-1
|
|
415951
|
Dithiothreitol, 99%
|
浜岀～铇囩硸閱�
|
3483-12-3
|
|
1914020
|
Phosphate Buffered Saline, 10×
|
PBS绶╂矕娑�
|
N/A
|
|
3910101
|
Protease Inhibitor Mix G
|
Cocktail铔嬬櫧閰舵姂鍒跺姂G
|
N/A
|
|
3910201
|
Protease Inhibitor Mix M
|
Cocktail铔嬬櫧閰舵姂鍒跺姂M
|
N/A
|
|
1201851
|
Protein Color Marker, M.W. 1,200 - 31,000 Da
|
铔嬬櫧闋愭煋Marker
|
N/A
|
|
1914019
|
SSC Buffer, 20×
|
SSC 绶╂矕娑�
|
N/A
|
|
1913953
|
Tris-Glycine-SDS Running Buffer, 10×
|
Tris-Glycine-SDS闆绘吵绶╂矕娑�
|
N/A
|
|
954773
|
Trypsin, 2500 units/mg
|
鑳拌泲鐧介叾
|
9002-07-7
|
鈻� 鍏朵粬閰嶅鐢�(ch菐n)鍝�锛�