Recognition of unique carboxyl-terminal motifs by distinct PDZ domains

Science

Volumn 275, Issue 5296, 1997, Pages 73-77

  Songyang, Z ^a   Fanning, A S ^a   Fu, C ^a   Xu, J ^a   Marfatia, S M ^a   Chishti, A H ^a   Crompton, A ^a   Chan, A C ^a   Andersen, J M ^a   Cantley, L C ^a  

^a NONE

Author keywords

[No Author keywords available]

Indexed keywords

CELL PROTEIN; N METHYL DEXTRO ASPARTIC ACID RECEPTOR;

ARTICLE; BINDING SITE; CARBOXY TERMINAL SEQUENCE; CRYSTAL STRUCTURE; HYDROPHILICITY; HYDROPHOBICITY; PRIORITY JOURNAL; PROTEIN BINDING; PROTEIN DOMAIN;

AMINO ACID SEQUENCE; ANIMALS; BINDING SITES; CRYSTALLOGRAPHY, X-RAY; GUANINE NUCLEOTIDE EXCHANGE FACTORS; GUANYLATE KINASE; HELMINTH PROTEINS; HUMANS; KINESIN; MEMBRANE PROTEINS; MODELS, MOLECULAR; MYOSINS; NERVE TISSUE PROTEINS; NUCLEOSIDE-PHOSPHATE KINASE; PEPTIDE LIBRARY; PEPTIDES; PROTEIN STRUCTURE, SECONDARY; PROTEIN STRUCTURE, TERTIARY; PROTEIN-TYROSINE-PHOSPHATASE; PROTEINS; SEQUENCE HOMOLOGY, AMINO ACID;

EID: 15644379801     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.275.5296.73     Document Type: Article

References (33)

1. G. B. Cohen, R. Ren, D. Baltimore, Cell 80, 237 (1995); T. Pawson, Nature 373, 573 (1995).
2. G. B. Cohen, R. Ren, D. Baltimore, Cell 80, 237 (1995); T. Pawson, Nature 373, 573 (1995).
3. D. F. Woods and P. J. Bryant, Cell 66, 451 (1991); Mech. Dev. 44, 85 (1993).
4. D. F. Woods and P. J. Bryant, Cell 66, 451 (1991); Mech. Dev. 44, 85 (1993).
5. K. O. Cho, C. A. Hunt, M. B. Kennedy, Neuron 9, 929 (1992); M. B. Kennedy, Trends Biochem. Sci. 20, 350 (1995).
6. K. O. Cho, C. A. Hunt, M. B. Kennedy, Neuron 9, 929 (1992); M. B. Kennedy, Trends Biochem. Sci. 20, 350 (1995).
7. M. Itoh et al., J. Cell Biol. 121, 491 (1993); E. Willott et al., Proc. Natl. Acad. Sci. U.S.A. 90, 7834 (1993).
8. M. Itoh et al., J. Cell Biol. 121, 491 (1993); E. Willott et al., Proc. Natl. Acad. Sci. U.S.A. 90, 7834 (1993).
9. C. P. Ponting and C. Phillips, Trends Biochem. Sci. 20, 102 (1995).
10. H. C. Kornau, L. T. Schenker, M. B. Kennedy, P. H. Seeburg, Science 269, 1737 (1995).
11. E. Kim, M. Niethammer, A. Rothschild, Y. N. Jan, M. Sheng, Nature 378, 85 (1995).
12. J. H. M. Cabral et al., ibid. 382, 649 (1996).
13. D. A. Doyle et al., Cell 85, 1067 (1996).
14. Z. Songyang et al., ibid. 72, 767 (1993).
15. A primary peptide library, KNXXXXXXXX-COOH, where X indicates all amino acids except Cys and Trp, was first used to screen peptides that bind specifically to the glutathione-S-transferase (GST)-PDZ domains. All the peptides in the library end with free carboxylate, therefore orienting all binding pockets. The peptides that bound were sequenced as a mixture, and the selectivities for amino acids at a given position were determined by comparison to the sequence of control experiments with GST alone (10). Arg was not included in the calculation because of buffer contamination during sequencing. A secondary library, KNXXXXXX(S,T,Y) XX-COOH, where the -2 position was fixed with Ser, Thr, and Tyr, was used to further define the preference of some PDZ domains.
16. Peptide library synthesis was as described (10). Individual PDZ domains were expressed and purified as GST fusion proteins: murine hDlg PDZ-1 (186-282), PDZ-2 (281-377), PDZ-3 (428-518), and PDZ-1/2 (281-518); murine PTPbas PDZ-3 (1351-1445) and PDZ-5 (1758-1848); murine Tiam-1 PDZ; human LIN-2 PDZ (422-507); human erythroid p55 PDZ (1-164); and human AF-6 PDZ (983-1102). Glutathione beads (50 to 60 μl) saturated with GST-PDZ proteins were mixed with the peptide library (1 mg) in 300 μl of TSN buffer [40 mM triethylamine (pH 7.6), 150 mM NaCl, and 0.01% NP-40] containing bovine serum albumin (BSA, 1 mg/ml) and 1 mM dithiothreitol (DTT). After 45 min of constant shaking at 4°C, the beads were washed with TSN buffer. The peptides retained were eluted with 30% acetic acid, lyophilized, resuspended in distilled water, and sequenced on a Bio-Applied 477A sequencer.
17. Z. Songyang and L. C. Cantley, unpublished data.
18. T. Sato, S. Irie, S. Kitada, J. C. Reed, Science 268, 411 (1995).
19. P. Ruff, D. W. Speicher, A. Husain-Chishti, Proc. Natl. Acad. Sci. U.S.A. 88, 6595 (1991).
20. R. Hoskins, A. F. Hajnal, S. A. Harp, S. K. Kim, Development 122, 97 (1996); A. Brecher et al., in preparation.
21. R. Hoskins, A. F. Hajnal, S. A. Harp, S. K. Kim, Development 122, 97 (1996); A. Brecher et al., in preparation.
22. G. G. Habets et al., Cell 77, 537 (1994).
23. R. Prasad et al., Cancer Res. 53, 5624 (1993).
24. G. Payne, S. E. Shoelson, G. D. Gish, T. Pawson, C. T. Walsh, Proc. Natl. Acad. Sci. U.S.A. 90, 4902 (1993).
25. H. T. Yu et al., Cell 76, 933 (1994).
26. Z. Songyang et al., data not shown.
27. J. S. Simske, S. M. Kaech, S. A. Harp, S. K. Kim, Cell 85, 195 (1996).
28. S. M. Marfatia et al., J. Biol. Chem., in press.
29. V. Hata, S. Butz, T. C. Sudhof, J. Neurosci. 16, 2488 (1996).
30. J. E. Brenman et al., Cell 84, 757 (1996).
31. B. Lipinska, M. Zylicz, C. Georgopoulos, J. Bacteriol. 172, 1791 (1990).
32. S. V. Shestakov et al., J. biol. Chem. 269, 19354 (1994).
33. We thank M. Berne for peptide synthesis and sequencing, R. Mackinnon for structural coordinates of PSD-95-3, W. Boll and A. Nguyen for technical assistance, A. Couvillon for antibodies to GST, M. Oishi and T. Woodford-Thomas for the PTPbas cDNA, and A. Brecher for human LIN-2 PDZ. C.F. is a Lucille Markey Fellow. Supported by grants from American Cancer Society and Lucille P. Markey Charitable Trust (L.C.C.), NIH grants CA66263 and DK34989 (J.M.A, and A.S.F), Pew Scholars Program (A.C.C.), and NIH grant CA66263 (A.H.C. and S.M.M.).