Reference: Honey S, et al. (2001)
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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.
- Reference Type
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Journal Article |
Research Support, U.S. Gov't, P.H.S.
- Authors
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Honey S,
Schneider BL,
Schieltz DM,
Yates JR,
Futcher B
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- SSA1 | CKS1 | CDC28 | CLB2 | SIC1 | CLB2-CDC28 kinase complex
Gene Ontology Annotations
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| Evidence ID |
Analyze ID |
Gene/Complex |
Systematic Name/Complex Accession |
Qualifier |
Gene Ontology Term ID |
Gene Ontology Term |
Aspect |
Annotation Extension |
Evidence |
Method |
Source |
Assigned On |
Reference |
Phenotype Annotations
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| Evidence ID |
Analyze ID |
Gene |
Gene Systematic Name |
Phenotype |
Experiment Type |
Experiment Type Category |
Mutant Information |
Strain Background |
Chemical |
Details |
Reference |
Disease Annotations
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| Evidence ID |
Analyze ID |
Gene |
Gene Systematic Name |
Disease Ontology Term |
Disease Ontology Term ID |
Qualifier |
Evidence |
Method |
Source |
Assigned On |
|
Reference |
Regulation Annotations
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| Evidence ID |
Analyze ID |
Regulator |
Regulator Systematic Name |
Target |
Target Systematic Name |
Direction |
Regulation of |
Happens During |
Regulator Type |
Direction |
Regulation Of |
Happens During |
Method |
Evidence |
Strain Background |
Reference |
Post-translational Modifications
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Site |
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Modification |
Modifier |
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Genetic Interactions
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| Evidence ID |
Analyze ID |
|
Interactor |
Interactor Systematic Name |
Interactor |
Interactor Systematic Name |
Allele |
Assay |
Annotation |
Action |
Phenotype |
SGA score |
P-value |
Source |
Reference |
Note |
Physical Interactions
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| Interactor | Interactor | Assay | Annotation | Action | Modification |
|---|
| CDC28 | CLB2 | Affinity Capture-MS | manually curated | Hit-Bait | No Modification |
| CKS1 | CLB2 | Affinity Capture-MS | manually curated | Hit-Bait | No Modification |
| CLB1 | CLB2 | Affinity Capture-MS | manually curated | Hit-Bait | No Modification |
| DBP1 | CLB2 | Affinity Capture-MS | manually curated | Hit-Bait | No Modification |
| DED1 | CLB2 | Affinity Capture-MS | manually curated | Hit-Bait | No Modification |
| PHO85 | CLB2 | Affinity Capture-MS | manually curated | Hit-Bait | No Modification |
| PSA1 | CLB2 | Affinity Capture-MS | manually curated | Hit-Bait | No Modification |
| SIC1 | CLB2 | Affinity Capture-MS | manually curated | Hit-Bait | No Modification |
| SLM1 | CLB2 | Affinity Capture-MS | manually curated | Hit-Bait | No Modification |
| SSA1 | CLB2 | Affinity Capture-MS | manually curated | Hit-Bait | No Modification |
Functional Complementation Annotations
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| Complement ID |
Locus ID |
Gene |
Species |
Gene ID |
Strain background |
Direction |
Details |
Source |
Reference |