A new monoclonal antibody enables BAR analysis of subcellular importin β1 interactomes

Didi-Andreas Song; Stefanie Alber; Ella Doron-Mandel; Vera Schmid; Christin A Albus; Orith Leitner; Hedva Hamawi; Juan A Oses-Prieto; Nili Dezorella; Alma L Burlingame; Mike Fainzilber; Ida Rishal

doi:10.1016/j.mcpro.2022.100418

A new monoclonal antibody enables BAR analysis of subcellular importin β1 interactomes

Didi-Andreas Song, Stefanie Alber, Ella Doron-Mandel, Vera Schmid, Christin A Albus, Orith Leitner, Hedva Hamawi, Juan A Oses-Prieto, Nili Dezorella, Alma L Burlingame, Mike Fainzilber, Ida Rishal

Weizmann Institute of Science

Research output: Contribution to journal › Article › peer-review

Abstract

Importin β1 (KPNB1) is a nucleocytoplasmic transport factor with critical roles in both cytoplasmic and nucleocytoplasmic transport, hence there is keen interest in the characterization of its subcellular interactomes. We found limited efficiency of BioID in detection of importin complex cargos, and therefore generated a highly specific and sensitive anti-KPNB1 monoclonal antibody to enable Biotinylation by Antibody Recognition (BAR) analysis of importin β1 interactomes. The monoclonal antibody recognizes an epitope comprising residues 301-320 of human KPBN1, and strikingly is highly specific for cytoplasmic KPNB1 in diverse applications, with little reaction with KPNB1 in the nucleus. BAR with this novel antibody revealed numerous new interactors of importin β1, expanding the KPNB1 interactome to cytoplasmic and signaling complexes that highlight potential new functions for the importins complex beyond nucleocytoplasmic transport. Data are available via ProteomeXchange with identifier PXD032728.

Original language	English
Article number	100418
Number of pages	15
Journal	Molecular & Cellular Proteomics
Volume	21
Issue number	11
Early online date	28 Sep 2022
DOIs	https://doi.org/10.1016/j.mcpro.2022.100418
State	Published - Nov 2022

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Song, D.-A., Alber, S., Doron-Mandel, E., Schmid, V., Albus, C. A., Leitner, O., Hamawi, H., Oses-Prieto, J. A., Dezorella, N., Burlingame, A. L., Fainzilber, M., & Rishal, I. (2022). A new monoclonal antibody enables BAR analysis of subcellular importin β1 interactomes. Molecular & Cellular Proteomics, 21(11), Article 100418. https://doi.org/10.1016/j.mcpro.2022.100418

@article{5ab189dd4096496a9b3b3dc1b1a14774,

title = "A new monoclonal antibody enables BAR analysis of subcellular importin β1 interactomes",

abstract = "Importin β1 (KPNB1) is a nucleocytoplasmic transport factor with critical roles in both cytoplasmic and nucleocytoplasmic transport, hence there is keen interest in the characterization of its subcellular interactomes. We found limited efficiency of BioID in detection of importin complex cargos, and therefore generated a highly specific and sensitive anti-KPNB1 monoclonal antibody to enable Biotinylation by Antibody Recognition (BAR) analysis of importin β1 interactomes. The monoclonal antibody recognizes an epitope comprising residues 301-320 of human KPBN1, and strikingly is highly specific for cytoplasmic KPNB1 in diverse applications, with little reaction with KPNB1 in the nucleus. BAR with this novel antibody revealed numerous new interactors of importin β1, expanding the KPNB1 interactome to cytoplasmic and signaling complexes that highlight potential new functions for the importins complex beyond nucleocytoplasmic transport. Data are available via ProteomeXchange with identifier PXD032728.",

author = "Didi-Andreas Song and Stefanie Alber and Ella Doron-Mandel and Vera Schmid and Albus, {Christin A} and Orith Leitner and Hedva Hamawi and Oses-Prieto, {Juan A} and Nili Dezorella and Burlingame, {Alma L} and Mike Fainzilber and Ida Rishal",

year = "2022",

month = nov,

doi = "10.1016/j.mcpro.2022.100418",

language = "الإنجليزيّة",

volume = "21",

journal = "Molecular & Cellular Proteomics",

issn = "1535-9476",

publisher = "American Society for Biochemistry and Molecular Biology Inc.",

number = "11",

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TY - JOUR

T1 - A new monoclonal antibody enables BAR analysis of subcellular importin β1 interactomes

AU - Song, Didi-Andreas

AU - Alber, Stefanie

AU - Doron-Mandel, Ella

AU - Schmid, Vera

AU - Albus, Christin A

AU - Leitner, Orith

AU - Hamawi, Hedva

AU - Oses-Prieto, Juan A

AU - Dezorella, Nili

AU - Burlingame, Alma L

AU - Fainzilber, Mike

AU - Rishal, Ida

PY - 2022/11

Y1 - 2022/11

N2 - Importin β1 (KPNB1) is a nucleocytoplasmic transport factor with critical roles in both cytoplasmic and nucleocytoplasmic transport, hence there is keen interest in the characterization of its subcellular interactomes. We found limited efficiency of BioID in detection of importin complex cargos, and therefore generated a highly specific and sensitive anti-KPNB1 monoclonal antibody to enable Biotinylation by Antibody Recognition (BAR) analysis of importin β1 interactomes. The monoclonal antibody recognizes an epitope comprising residues 301-320 of human KPBN1, and strikingly is highly specific for cytoplasmic KPNB1 in diverse applications, with little reaction with KPNB1 in the nucleus. BAR with this novel antibody revealed numerous new interactors of importin β1, expanding the KPNB1 interactome to cytoplasmic and signaling complexes that highlight potential new functions for the importins complex beyond nucleocytoplasmic transport. Data are available via ProteomeXchange with identifier PXD032728.

AB - Importin β1 (KPNB1) is a nucleocytoplasmic transport factor with critical roles in both cytoplasmic and nucleocytoplasmic transport, hence there is keen interest in the characterization of its subcellular interactomes. We found limited efficiency of BioID in detection of importin complex cargos, and therefore generated a highly specific and sensitive anti-KPNB1 monoclonal antibody to enable Biotinylation by Antibody Recognition (BAR) analysis of importin β1 interactomes. The monoclonal antibody recognizes an epitope comprising residues 301-320 of human KPBN1, and strikingly is highly specific for cytoplasmic KPNB1 in diverse applications, with little reaction with KPNB1 in the nucleus. BAR with this novel antibody revealed numerous new interactors of importin β1, expanding the KPNB1 interactome to cytoplasmic and signaling complexes that highlight potential new functions for the importins complex beyond nucleocytoplasmic transport. Data are available via ProteomeXchange with identifier PXD032728.

UR - http://www.scopus.com/inward/record.url?scp=85141330024&partnerID=8YFLogxK

U2 - 10.1016/j.mcpro.2022.100418

DO - 10.1016/j.mcpro.2022.100418

M3 - مقالة

SN - 1535-9476

VL - 21

JO - Molecular & Cellular Proteomics

JF - Molecular & Cellular Proteomics

IS - 11

M1 - 100418

ER -

A new monoclonal antibody enables BAR analysis of subcellular importin β1 interactomes

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