title: AFAP-120. A variant form of the Src SH2/SH3-binding partner AFAP-110 is detected in brain and contains a novel internal sequence which binds to a 67-kDa protein.
abstract: SH2 and SH3 domains have been characterized as functional domains that mediate protein-protein interactions in signal transduction. Recently, the cDNA sequence of a novel Src- and Fyn-binding protein called AFAP-110, for Actin-Filament Associated Protein-110 kDa, was reported. This protein was distinctive in that it is both an SH2 and SH3 binding partner for the non-receptor tyrosine kinases Src and Fyn. Here, we report the characterization of an alternatively processed form of AFAP-110 that encodes an additional 258 base pair (bp) of open reading frame. Transient expression of this full-length clone reveals a molecular mass of 120 kDa. Western blot analysis indicate that a larger 120-kDa variant of AFAP-110 can be detected in brain and is not detectable in any other tissues examined. Northern blot analysis indicate that the novel 258-bp insert can be detected in brain RNA but not chick embryo fibroblast RNA. We propose the name AFAP-120, for Actin Filament-Associated Protein-120 kDa. Expression of the 258-bp novel insert (NINS) as a glutathione S-transferase-encoded fusion protein permits adsorption of a 67-kDa protein from tissue lysates. Deletion analysis of the NINS indicates that the interaction with p67 can be attributed to a proline-rich motif that resembles an SH3-binding motif. We hypothesize that AFAP-120 facilitates interactions in brain between SH2/SH3 signaling proteins and actin filaments and that a proline-rich motif in the NINS may exist to facilitate additional interactions between cellular proteins in brain and actin filaments.
authors.affiliation: Mary Babb Randolph Cancer Center, Morgantown, West Virginia.
authors.forename: D C
authors.initials: DC
authors.lastname: Flynn
authors.forename: T C
authors.initials: TC
authors.lastname: Koay
authors.forename: C G
authors.initials: CG
authors.lastname: Humphries
authors.forename: A C
authors.initials: AC
authors.lastname: Guappone
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D001921:Brain / Q000378:metabolism*; D002460:Cell Line; D002478:Cells, Cultured; D002642:Chick Embryo; D003001:Cloning, Molecular; D018076:DNA, Complementary; D008840:Microfilament Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding
chemical_list: C084510:AFAP 110; D018076:DNA, Complementary; D008840:Microfilament Proteins; D010750:Phosphoproteins