PMID:11950598

Citation	Paris, S, Za, L, Sporchia, B and de Curtis, I (2002) Analysis of the subcellular distribution of avian p95-APP2, an ARF-GAP orthologous to mammalian paxillin kinase linker. Int. J. Biochem. Cell Biol. 34:826-37
Abstract	We describe here the identification and characterization of avian p95-APP2, a multi-domain protein of a recently identified family of ADP-ribosylation factor (ARF)-GTPase-activating proteins (GAPs) including mammalian G protein-coupled receptor kinases (GRK)-interactor 1 (GIT1), paxillin kinase linker (PKL), and GIT2, as well as avian p95-APP1. The p95-APP2 is eluted from Rac-GTP-gamma-S, but not from Rac-GDP-beta-S columns. As other members of the family, p95-APP2 has binding regions for the focal adhesion protein paxillin, and for the Rac exchanging factor PIX. Sequence comparison indicates that p95-APP2 is the avian orthologue of mammalian PKL. Expression studies showed a largely diffuse distribution of the full length p95-APP2, without evident effects on cell morphology. We observed a dramatic difference between the localization of the amino-terminal portion of the protein, including the ARF-GAP domain and the three ankyrin repeats, and the carboxy-terminal portion including the paxillin-binding site. Moreover, the expression of truncated carboxy-terminal polypeptides including both the PIX- and paxillin-binding regions leads to a marked localization of the protein together with paxillin at large vesicles. Comparison of the expression of corresponding ARF-GAP-deficient constructs from p95-APP2 and p95-APP1 shows their distribution at distinct endocytic compartments. Altogether, these data support a role of distinct members of this family of ARF-GAPs in the regulation of different steps of membrane traffic during cell motility, and suggest that p95-APP2 may shuttle between an intracellular compartment and the cell periphery, although, further work will be needed to address this point.
Links	PubMed
Keywords	ADP-Ribosylation Factors/chemistry; ADP-Ribosylation Factors/genetics; ADP-Ribosylation Factors/metabolism; Adaptor Proteins, Signal Transducing; Amino Acid Sequence; Animals; Base Sequence; Carrier Proteins/chemistry; Carrier Proteins/genetics; Carrier Proteins/metabolism; Cell Cycle Proteins; Cell Movement/physiology; Cells, Cultured; Chick Embryo; Chickens; Cloning, Molecular; DNA, Complementary/genetics; Fibroblasts/metabolism; GTPase-Activating Proteins/chemistry; GTPase-Activating Proteins/genetics; GTPase-Activating Proteins/metabolism; Humans; Macromolecular Substances; Molecular Sequence Data; Phosphoproteins; Protein Structure, Tertiary; Sequence Deletion; Species Specificity; Subcellular Fractions/metabolism
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
RAT:ARHG7	contributes_to	GO:0031267: small GTPase binding	ECO:0000353: physical interaction evidence used in manual assertion	UniProtKB:Q13153,UniProtKB:Q9DE64	F	has_input:(UniProtKB:O93466)	Seeded From UniProt	complete
RAT:ARHG7	part_of	GO:0032991: protein-containing complex	ECO:0000314: direct assay evidence used in manual assertion		C		Seeded From UniProt	complete
HUMAN:PAK1	contributes_to	GO:0031267: small GTPase binding	ECO:0000353: physical interaction evidence used in manual assertion	UniProtKB:O55043,UniProtKB:Q9DE64	F	has_input:(UniProtKB:O93466)	Seeded From UniProt	complete
CHICK:Q9DE64	contributes_to	GO:0031267: small GTPase binding	ECO:0000353: physical interaction evidence used in manual assertion	UniProtKB:O55043,UniProtKB:Q13153	F	has_input:(UniProtKB:O93466)	Seeded From UniProt	complete
CHICK:O93466	enables	GO:0044877: protein-containing complex binding	ECO:0000314: direct assay evidence used in manual assertion		F	has_input:(UniProtKB:O55043) has_input:(UniProtKB:Q13153) has_input:(UniProtKB:Q9DE64)	Seeded From UniProt	complete
CHICK:Q9DE64	contributes_to	GO:0048365: Rac GTPase binding	ECO:0000353: physical interaction evidence used in manual assertion	UniProtKB:O55043,UniProtKB:Q13153	F	has_input:(UniProtKB:O93466)	Seeded From UniProt	complete
CHICK:Q9DE64	part_of	GO:0032991: protein-containing complex	ECO:0000314: direct assay evidence used in manual assertion		C		Seeded From UniProt	complete
HUMAN:PAK1	part_of	GO:0032991: protein-containing complex	ECO:0000314: direct assay evidence used in manual assertion		C		Seeded From UniProt	complete
HUMAN:PAK1	contributes_to	GO:0048365: Rac GTPase binding	ECO:0000353: physical interaction evidence used in manual assertion	UniProtKB:O55043,UniProtKB:Q9DE64	F	has_input:(UniProtKB:O93466)	Seeded From UniProt	complete

References

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