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PMID:16275660

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Citation

Grelle, G, Kostka, S, Otto, A, Kersten, B, Genser, KF, Müller, EC, Wälter, S, Böddrich, A, Stelzl, U, Hänig, C, Volkmer-Engert, R, Landgraf, C, Alberti, S, Höhfeld, J, Strödicke, M and Wanker, EE (2006) Identification of VCP/p97, carboxyl terminus of Hsp70-interacting protein (CHIP), and amphiphysin II interaction partners using membrane-based human proteome arrays. Mol. Cell Proteomics 5:234-44

Abstract

Proteins mediate their biological function through interactions with other proteins. Therefore, the systematic identification and characterization of protein-protein interactions have become a powerful proteomic strategy to understand protein function and comprehensive cellular regulatory networks. For the screening of valosin-containing protein, carboxyl terminus of Hsp70-interacting protein (CHIP), and amphiphysin II interaction partners, we utilized a membrane-based array technology that allows the identification of human protein-protein interactions with crude bacterial cell extracts. Many novel interaction pairs such as valosin-containing protein/autocrine motility factor receptor, CHIP/caytaxin, or amphiphysin II/DLP4 were identified and subsequently confirmed by pull-down, two-hybrid and co-immunoprecipitation experiments. In addition, assays were performed to validate the interactions functionally. CHIP e.g. was found to efficiently polyubiquitinate caytaxin in vitro, suggesting that it might influence caytaxin degradation in vivo. Using peptide arrays, we also identified the binding motifs in the proteins DLP4, XRCC4, and fructose-1,6-bisphosphatase, which are crucial for the association with the Src homology 3 domain of amphiphysin II. Together these studies indicate that our human proteome array technology permits the identification of protein-protein interactions that are functionally involved in neurodegenerative disease processes, the degradation of protein substrates, and the transport of membrane vesicles.

Links

PubMed Online version:10.1074/mcp.M500198-MCP200

Keywords

Adenosine Triphosphatases; Amino Acid Sequence; Animals; COS Cells; Cell Cycle Proteins/chemistry; Cell Cycle Proteins/metabolism; Cercopithecus aethiops; HSC70 Heat-Shock Proteins/metabolism; Humans; Membranes, Artificial; Molecular Sequence Data; Nerve Tissue Proteins/metabolism; Protein Array Analysis; Protein Binding; Protein Interaction Mapping; Protein Structure, Tertiary; Proteome

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

HUMAN:BIN1

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P09467

F

Seeded From UniProt

complete

HUMAN:BIN1

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q13426

F

Seeded From UniProt

complete

HUMAN:BIN1

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q9Y2H0

F

Seeded From UniProt

complete

HUMAN:HS90A

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q9UNE7

F

Seeded From UniProt

complete

HUMAN:F16P1

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:O00499

F

Seeded From UniProt

complete

HUMAN:TAU

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q9UNE7

F

Seeded From UniProt

complete

HUMAN:HSP7C

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q9UNE7

F

Seeded From UniProt

complete

HUMAN:TERA

located_in

GO:0048471: perinuclear region of cytoplasm

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:TERA

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q9UKV5

F

Seeded From UniProt

complete

HUMAN:TERA

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q9UNZ2

F

Seeded From UniProt

complete

HUMAN:XRCC4

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:O00499

F

Seeded From UniProt

complete

HUMAN:UB2Q1

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q9UNE7

F

Seeded From UniProt

complete

HUMAN:ATCAY

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q9UNE7

F

Seeded From UniProt

complete

HUMAN:AMFR

located_in

GO:0048471: perinuclear region of cytoplasm

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:AMFR

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P55072

F

Seeded From UniProt

complete

HUMAN:CHIP

involved_in

GO:0000209: protein polyubiquitination

ECO:0000314: direct assay evidence used in manual assertion

P

  • has_input:(UniProtKB:Q86WG3)

Seeded From UniProt

complete

HUMAN:CHIP

part_of

GO:0000151: ubiquitin ligase complex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:CHIP

enables

GO:0061630: ubiquitin protein ligase activity

ECO:0000314: direct assay evidence used in manual assertion

F

  • has_input:(UniProtKB:Q86WG3)

Seeded From UniProt

complete

HUMAN:CHIP

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P07900

F

Seeded From UniProt

complete

HUMAN:CHIP

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P10636

F

Seeded From UniProt

complete

HUMAN:CHIP

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P11142

F

Seeded From UniProt

complete

HUMAN:CHIP

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q7Z7E8

F

Seeded From UniProt

complete

HUMAN:CHIP

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q86WG3

F

Seeded From UniProt

complete

HUMAN:NSF1C

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P55072

F

Seeded From UniProt

complete

HUMAN:DLGP4

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:O00499

F

Seeded From UniProt

complete

HUMAN:TERA

part_of

GO:0048471: perinuclear region of cytoplasm

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:HSP71

GO:0005515: protein binding

IPI: Inferred from Physical Interaction: UniProtKB:Q9UNE7

F

Seeded From UniProt

complete



See also

References

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