Characterization of the Interaction of Natural Proline-Rich Peptides with Five Different SH3 Domains

Viguera, Ana Rosa; Arrondo, José Luis R.; Musacchio, Andrea; Saraste, Matti; Serrano, Luís

doi:10.1021/bi00202a011

Cited by 138 publications

(145 citation statements)

References 43 publications

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“…Third, the homology between DOC-2 and p96/mDab2 is weakened towards the carboxy-terminus of the protein. They both have proline-rich domains in this region which contains multiple SH3 binding motifs Feng et al, 1994;Sparks et al, 1994;Viguera et al, 1994). Since Xu et al (1997) has recently demonstrated that the proline-rich domain of mouse p96/mDab2 can interact with the SH3 domains of Grb2 and reduce the binding between Grb2 and Sos, it will be interesting to examine whether the proline-rich domain of DOC-2 can also interact with these signal transduction molecules in the HOSE cells.…”

Section: Discussionmentioning

confidence: 99%

DOC-2, a candidate tumor suppressor gene in human epithelial ovarian cancer

et al. 1998

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Using RNA ®ngerprinting (RAP) strategy and Northern blot analysis, we identi®ed a di erentially expressed sequence DOC-2 which is detectable in all normal human ovarian surface epithelial (HOSE) cell cultures but not in ovarian cancer cell lines and tissues. Subsequent cloning of DOC-2 from a cDNA library generated from the HOSE cells was carried out using the 3' and 5' RACE approach. A 3268 base pair full length cDNA of DOC-2 was isolated and sequenced. The predicted protein has a length of 770 amino acids. Homology search of all NCBI sequences indicated that the amino acid sequence of DOC-2 shares 93% homology with the mouse p96/ mDab2 phosphoprotein and has a phosphotyrosine interacting domain (PID) and multiple SH3 binding motifs. Chromosomal localization by FISH showed that the DOC-2 gene is located on 5p13. Western blot analysis showed that the 105 kDa DOC-2 protein was down-regulated in all the carcinoma cell lines. In-situ immunohistochemistry performed on normal ovaries, and benign, borderline and invasive ovarian tumor tissues showed down regulation of DOC-2 protein particularly in serous ovarian tumor tissues. When DOC-2 was transfected into the ovarian carcinoma cell line SKOV3, the stable transfectants showed signi®cantly reduced growth rate and ability to form tumors in nude mice. These data suggest that down-regulation of DOC-2 may play an important role in ovarian carcinogenesis.

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Section: Discussionmentioning

confidence: 99%

DOC-2, a candidate tumor suppressor gene in human epithelial ovarian cancer

et al. 1998

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“…To evaluate the affinity and selectivity of C⌬4, the high-affinity 11-residue peptide p85␣-2 and the lowaffinity 13-residue peptide p85␣-1 were selected as target ligands (15). Both peptides were obtained commercially (United Biochemical Research, Seattle) and used without further purification.…”

Section: Methodsmentioning

confidence: 99%

Engineering a signal transduction mechanism for protein-based biosensors

Kohn

Plaxco

2005

Proc. Natl. Acad. Sci. U.S.A.

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Hybridization-induced conformational changes have been successfully used in biosensors for the transduction of DNA-binding events into readily observable optical or electronic signals. Similar signal transduction has not, however, proven of equal utility in proteinbased biosensors. The discrepancy arises because, unlike ssDNA, most proteins do not undergo significant conformational changes upon ligand binding. Here, we describe a solution to this problem. We show that an arbitrarily selected, normally well folded protein can be rationally engineered such that it undergoes ligand-induced folding. The engineered protein responds rapidly (milliseconds) and selectively to its target, and it couples recognition with the largest possible conformational change: folding. These traits suggest that ligand-induced folding could serve as an ideal signaltransduction mechanism. Consistent with this claim, we demonstrate a label-free optical biosensor based on the effect that is sufficiently selective to detect its target even in complex, contaminant-ridden samples such as blood serum.fluorescence ͉ molecular beacon ͉ natively unfolded ͉ quenching

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“…3). In contrast, the latter spectral transition gives rise to a weakly positive band at 225 nm for Ac-RALPPLPRY-NH2, indicative of polyproline II helical conformation with trans Xaa-Pro bonds [27][28][29][30][31][32]. Similarly, Fig.…”

Section: Structural Properties Of Peptides 1--5mentioning

confidence: 98%

Extended binding sites of cyclophilin as revealed by the interaction with HIV‐1 Gag polyprotein derived oligopeptides

et al. 1996

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Oligopeptides derived from the gag polyprotein (Pr55 gag) of human immunodeficiency virus type 1 (HIV-1) segment were used to evaluate the extension of the putative binding region for the complex of Pr55 gag and the human cytosolic peptidyl prolyl cis/trans isomerase (PPIase) 18 kDa cyclophilin (Cypl8). Five N-terminally acetylated, C-terminally amidated oligopeptides containing one (HIV-1 Gag218-224; 1), two (HIV-1 Gag 21s-226 and HIV-1 Gag217-224; 2 and 3, respectively), three (HIV-I Gag217-226; 4) or four (HIV-1 Gag213-237; 5) proline residues were synthesized. Using competition experiments with a standard substrate the binding affinities to Cypl8 of the synthesized peptides were determined. The ICs0 value of 184 ~M for the 25-mer peptide 5 was fivefold or more lower than those of the peptides 1--4 lacking one or more prolines. Failure of competition in assays containing enzymes of other PPlase families by millimolar concentrations of 5 revealed a Cyp18 specific interaction involving the active site of the enzyme. In its far UV circular dichroism, aqueous solutions of 5 display properties of random coil conformation, but spectra were also consistent with a small contribution of proline specific secondary structures. However, a proline-rich peptide typical of forming left-handed polyproline II helices did not compete for the active site of Cypl8. The results demonstrate that the putative binding region of HIV-1 gag polyprotein has a certain degree of binding affinity to the PPIase site of Cypl8, and may add a previously unrecognized topological component to the known subsite specificity of cyciophilins.

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Characterization of the Interaction of Natural Proline-Rich Peptides with Five Different SH3 Domains

Cited by 138 publications

References 43 publications

DOC-2, a candidate tumor suppressor gene in human epithelial ovarian cancer

DOC-2, a candidate tumor suppressor gene in human epithelial ovarian cancer

Engineering a signal transduction mechanism for protein-based biosensors

Extended binding sites of cyclophilin as revealed by the interaction with HIV‐1 Gag polyprotein derived oligopeptides

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