A novel multiple affinity purification tag and its use in identification of proteins associated with a cyclin-CDK complex

Sangeet Honey; Brandt L. Schneider; David M. Schieltz; John R. Yates; Bruce Futcher

doi:10.1093/nar/29.4.e24

A novel multiple affinity purification tag and its use in identification of proteins associated with a cyclin-CDK complex

Sangeet Honey
, Brandt L. Schneider
, David M. Schieltz
, John R. Yates
, Bruce Futcher

Microbiology and Immunology

Cold Spring Harbor Laboratory
University of Washington

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

10 Scopus citations

Abstract

A novel multiple affinity purification (MAFT) or tandem affinity purification (TAP) tag has been constructed. It consists of the calmodulin binding peptide, six histidine residues, and three copies of the hemagglutinin epitope. This 'CHH' MAFT tag allows two or three consecutive purification steps, giving high purity. Active Clb2-Cdc28 kinase complex was purified from yeast cells after inserting the CHH tag into Clb2. Associated proteins were identified using mass spectrometry. These included the known associated proteins Cdc28, Sic1 and Cks1. Several other proteins were found including the 70 kDa chaperone, Ssa1.

Original language	English
Article number	e24
Journal	Nucleic Acids Research
Volume	29
Issue number	4
DOIs	https://doi.org/10.1093/nar/29.4.e24
State	Published - Feb 1 2021

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10.1093/nar/29.4.e24

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abstract = "A novel multiple affinity purification (MAFT) or tandem affinity purification (TAP) tag has been constructed. It consists of the calmodulin binding peptide, six histidine residues, and three copies of the hemagglutinin epitope. This 'CHH' MAFT tag allows two or three consecutive purification steps, giving high purity. Active Clb2-Cdc28 kinase complex was purified from yeast cells after inserting the CHH tag into Clb2. Associated proteins were identified using mass spectrometry. These included the known associated proteins Cdc28, Sic1 and Cks1. Several other proteins were found including the 70 kDa chaperone, Ssa1.",

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AU - Honey, Sangeet

AU - Schneider, Brandt L.

AU - Schieltz, David M.

AU - Yates, John R.

AU - Futcher, Bruce

PY - 2021/2/1

Y1 - 2021/2/1

N2 - A novel multiple affinity purification (MAFT) or tandem affinity purification (TAP) tag has been constructed. It consists of the calmodulin binding peptide, six histidine residues, and three copies of the hemagglutinin epitope. This 'CHH' MAFT tag allows two or three consecutive purification steps, giving high purity. Active Clb2-Cdc28 kinase complex was purified from yeast cells after inserting the CHH tag into Clb2. Associated proteins were identified using mass spectrometry. These included the known associated proteins Cdc28, Sic1 and Cks1. Several other proteins were found including the 70 kDa chaperone, Ssa1.

AB - A novel multiple affinity purification (MAFT) or tandem affinity purification (TAP) tag has been constructed. It consists of the calmodulin binding peptide, six histidine residues, and three copies of the hemagglutinin epitope. This 'CHH' MAFT tag allows two or three consecutive purification steps, giving high purity. Active Clb2-Cdc28 kinase complex was purified from yeast cells after inserting the CHH tag into Clb2. Associated proteins were identified using mass spectrometry. These included the known associated proteins Cdc28, Sic1 and Cks1. Several other proteins were found including the 70 kDa chaperone, Ssa1.

UR - https://www.scopus.com/pages/publications/0035866279

U2 - 10.1093/nar/29.4.e24

DO - 10.1093/nar/29.4.e24

M3 - Article

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AN - SCOPUS:0035866279

SN - 0305-1048

VL - 29

JO - Nucleic Acids Research

JF - Nucleic Acids Research

IS - 4

M1 - e24

ER -

A novel multiple affinity purification tag and its use in identification of proteins associated with a cyclin-CDK complex

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