Differential affinity purification and mass spectrometry analysis of two nuclear pore complex isoforms in yeast S. cerevisiae

被引：0

作者：

Bensidoun, Pierre ^{[1
,2
,4
]}

Zenklusen, Daniel ^{[2
]}

Oeffinger, Marlene ^{[1
,2
,3
]}

机构：

[1] Inst Rech Clin Montreal, 110 Ave Pins Ouest, Montreal, PQ H2W 1R7, Canada

[2] Univ Montreal, Dept Biochim & Med Mol, Fac Med, Montreal, PQ H3T 1J4, Canada

[3] McGill Univ, Fac Med, Div Expt Med, Montreal, PQ H3A 1A3, Canada

[4] Inst Genet Mol Montpellier IGMM, 1919 Route Mende, F-34090 Montpellier, France

来源：

STAR PROTOCOLS | 2023年 / 4卷 / 03期

基金：

加拿大自然科学与工程研究理事会;

关键词：

PROTEIN; MLP2P;

D O I：

10.1016/j.xpro.2023.102359

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Two isoforms of the nuclear pore complex (NPC) have been identified in the yeast S. cerevisiae, which coexist at the periphery of the nucleus and differ by the presence or absence of a nuclear basket. Here, we present a protocol to isolate the two types of NPCs from the same cell extract and dissect their interactomes. We describe steps for powder preparation and magnetic bead conjunction and detail differential affinity purification and outcome evaluation through SDSPAGE, silver staining, and mass spectrometry analysis. For complete details on the use and execution of this protocol, please refer to Bensidoun et al.1

引用

页数：21

共 14 条

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JOURNAL OF CELL BIOLOGY, 2012, 196 (01): : 19 - 27
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DATA IN BRIEF, 2015, 2 : 29 - 31
[3] Analysis of Chloroquine Resistance Transporter (CRT) Isoforms and Orthologues in S. cerevisiae Yeast
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BIOCHEMISTRY, 2011, 50 (31) : 6701 - 6710
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JOURNAL OF CELL BIOLOGY, 2015, 208 (06): : 729 - 744
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MOLECULAR BIOLOGY OF THE CELL, 2000, 11 : 151A - 151A
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Kralt, Annemarie
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Agote-Aran, Arantxa
Mancini, Roberta
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ELIFE, 2022, 11
[7] ANALYSIS OF PIPERAQUINE TRANSPORT BY PLASMODIUM FALCIPARUM CHLOROQUINE RESISTANCE TRANSPORTER ISOFORMS HETEROLOGOUSLY EXPRESSED IN S. CEREVISIAE YEAST
Riegel, Bryce E.
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AMERICAN JOURNAL OF TROPICAL MEDICINE AND HYGIENE, 2019, 101 : 274 - 274
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ANALYTICAL CHEMISTRY, 2020, 92 (02) : 1874 - 1882
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Luz Valero, M.
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DATA IN BRIEF, 2016, 6 : 965 - 969

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