Anal Biochem 2011,
PMID: 21146485
Park, Miki Susanto; Chu, Feixia; Xie, Jinghang; Wang, Yu; Bhattacharya, Pompeya; Chan, William K
Cyclophilin-40 (CyP40) is part of the immunophilin family and is found in Hsp90-containing protein complexes. We were interested in identifying proteins that interact with CyP40. CyP40-interacting proteins in HeLa cells were identified using the tandem affinity purification approach. Adenovirus expressing human CyP40 protein (Ad-CyP40), fused with streptavidin and calmodulin binding peptides at the N terminus, was generated. Proteins were separated on a sodium dodecyl sulfate-polyacrylamide gel electrophoresis gel after tandem affinity purification. Here 10 silver-stained protein bands that were enriched in the Ad-CyP40-infected lysate and the corresponding regions in the control lysate were excised, digested by trypsin, and identified by tandem mass spectrometric analysis. Of 11 interacting proteins that were identified, 4 (RACK1, Ku70, RPS3, and NF45) were expressed in rabbit reticulocyte lysate, bacteria, and MCF-7 cells. We confirmed that these proteins interact with CyP40. We observed that RACK1 suppressed the cobalt chloride-induced, hypoxia response element-dependent luciferase activity in MCF-7 cells but not in MCF-7 stable cells expressing approximately 10% of the cellular CyP40 content. In addition, RACK1 reduced the HIF-1α protein accumulation after cobalt chloride treatment, which was not observed when the CyP40 content was down-regulated. Collectively, we conclude that reduction of the HIF-1 α protein by RACK1 is CyP40-mediated.
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Text Mining Data
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Manually curated Databases
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IRef Biogrid Interaction:
PPID
—
HSP90AA1
(association, biochemical)
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IRef Biogrid Interaction:
XRCC6
—
PPID
(association, biochemical)
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IRef Biogrid Interaction:
XRCC6
—
PPID
(physical association, affinity chromatography technology)
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IRef Biogrid Interaction:
GNB2L1
—
PPID
(physical association, affinity chromatography technology)
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IRef Biogrid Interaction:
GNB2L1
—
PPID
(association, biochemical)
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IRef Biogrid Interaction:
PPID
—
RPS3
(association, biochemical)
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IRef Biogrid Interaction:
PPID
—
RPS3
(physical association, affinity chromatography technology)
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IRef Biogrid Interaction:
ILF2
—
PPID
(physical association, affinity chromatography technology)
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IRef Biogrid Interaction:
ILF2
—
PPID
(association, biochemical)
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IRef Biogrid Interaction:
TUBA1A
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PPID
(association, biochemical)
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IRef Biogrid Interaction:
HSPA1A
—
PPID
(association, biochemical)
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IRef Biogrid Interaction:
TUBB
—
PPID
(association, biochemical)
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IRef Biogrid Interaction:
PPID
—
HSP90AB1
(association, biochemical)
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IRef Intact Interaction:
XRCC6
—
PPID
(direct interaction, anti tag coimmunoprecipitation)
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IRef Intact Interaction:
XRCC6
—
PPID
(physical association, anti tag coimmunoprecipitation)
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IRef Intact Interaction:
GNB2L1
—
PPID
(physical association, anti tag coimmunoprecipitation)
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IRef Intact Interaction:
GNB2L1
—
PPID
(direct interaction, anti tag coimmunoprecipitation)
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IRef Intact Interaction:
PPID
—
RPS3
(physical association, anti tag coimmunoprecipitation)
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IRef Intact Interaction:
PPID
—
RPS3
(direct interaction, anti tag coimmunoprecipitation)
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IRef Intact Interaction:
ILF2
—
PPID
(physical association, anti tag coimmunoprecipitation)
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IRef Intact Interaction:
ILF2
—
PPID
(direct interaction, anti tag coimmunoprecipitation)
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IRef Intact Interaction:
Complex of 12 proteins
(association, tandem affinity purification)
In total, 58 gene pairs are associated to this article in curated databases