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Volumn 278, Issue 5340, 1997, Pages 1125-1128

Structural plasticity in a remodeled protein-protein interface

Author keywords

[No Author keywords available]

Indexed keywords

ARTICLE; BINDING AFFINITY; GENE DELETION; GENE REARRANGEMENT; NERVE CELL PLASTICITY; PRIORITY JOURNAL; PROTEIN DOMAIN; PROTEIN PROTEIN INTERACTION; PROTEIN STRUCTURE; STRUCTURE ANALYSIS;

EID: 0030707656     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.278.5340.1125     Document Type: Article
Times cited : (167)

References (35)
  • 1
    • 0025123333 scopus 로고
    • J. Janin and C. Chothia, J. Biol. Chem. 265, 16027 (1990); S. Jones and J. Thornton, Proc. Natl. Acad. Sci. U.S.A. 93, 13 (1996).
    • (1990) J. Biol. Chem. , vol.265 , pp. 16027
    • Janin, J.1    Chothia, C.2
  • 4
    • 0026598960 scopus 로고    scopus 로고
    • A. M. De Vos, M. Ultsch, A. A. Kossiakoff, Science 255, 306 (1992); M. Sundstrom et al., J. Biol. Chem. 271, 32197 (1996). The wild-type complex coordinates used in our study are from a 1:1 complex between G120R-hGH and hGHbp determined at 2.6 A resolution (T. Clackson, M. Ultsch, J. Wells, A. M. De Vos, in preparation).
    • (1992) Science , vol.255 , pp. 306
    • De Vos, A.M.1    Ultsch, M.2    Kossiakoff, A.A.3
  • 5
    • 0029770041 scopus 로고    scopus 로고
    • A. M. De Vos, M. Ultsch, A. A. Kossiakoff, Science 255, 306 (1992); M. Sundstrom et al., J. Biol. Chem. 271, 32197 (1996). The wild-type complex coordinates used in our study are from a 1:1 complex between G120R-hGH and hGHbp determined at 2.6 A resolution (T. Clackson, M. Ultsch, J. Wells, A. M. De Vos, in preparation).
    • (1996) J. Biol. Chem. , vol.271 , pp. 32197
    • Sundstrom, M.1
  • 6
    • 0026598960 scopus 로고    scopus 로고
    • in preparation
    • A. M. De Vos, M. Ultsch, A. A. Kossiakoff, Science 255, 306 (1992); M. Sundstrom et al., J. Biol. Chem. 271, 32197 (1996). The wild-type complex coordinates used in our study are from a 1:1 complex between G120R-hGH and hGHbp determined at 2.6 A resolution (T. Clackson, M. Ultsch, J. Wells, A. M. De Vos, in preparation).
    • Clackson, T.1    Ultsch, M.2    Wells, J.3    De Vos, A.M.4
  • 8
    • 0027132013 scopus 로고
    • B. C. Cunningham and J. A. Wells, Science 244, 1081 (1989); J. Mol. Biol. 234, 554 (1993).
    • (1993) J. Mol. Biol. , vol.234 , pp. 554
  • 10
    • 0028916599 scopus 로고
    • S. H. Bass, M. G. Mulkerrin, J. A. Wells, Proc. Natl. Acad. Sci. U.S.A. 88, 4498 (1991); T. Clackson and J. A. Wells, Science 267, 383 (1995).
    • (1995) Science , vol.267 , pp. 383
    • Clackson, T.1    Wells, J.A.2
  • 11
    • 1842416139 scopus 로고    scopus 로고
    • note
    • The single letter abbreviations for amino acid residues are as follows: A, Ala; C, Cys; D, Asp; E, Glu; F, Phe; G, Gly; H, His; I, Ile; K, Lys; L, Leu; M, Met; N, Asn; P, Pro; Q, Gln; R, Arg; S, Ser; T, Thr; V, Val; W, Trp; and Y, Tyr. Mutants are identified by the wildtype residue, followed by its position, and the mutant residue. Multiple mutants are indicated by a series of single mutants separated by commas.
  • 12
    • 0025888264 scopus 로고
    • 7. The plasmid pH0753 was derived from phGHam-g3 [H. B. Lowman, S. H. Bass, N. Simpson, J. A. Wells, Biochemistry 30, 10832 (1991)] by mutation of the base at position 419 from C to G in order to eliminate an Xba I restriction site.
    • (1991) Methods Enzymol. , vol.204 , pp. 125
    • Kunkel, T.A.1    Bebenek, K.2    McClary, J.3
  • 13
    • 0026343486 scopus 로고
    • 7. The plasmid pH0753 was derived from phGHam-g3 [H. B. Lowman, S. H. Bass, N. Simpson, J. A. Wells, Biochemistry 30, 10832 (1991)] by mutation of the base at position 419 from C to G in order to eliminate an Xba I restriction site.
    • (1991) Biochemistry , vol.30 , pp. 10832
    • Lowman, H.B.1    Bass, S.H.2    Simpson, N.3    Wells, J.A.4
  • 14
    • 1842298302 scopus 로고    scopus 로고
    • note
    • Phage displaying mutants of hGH were selected for binding to W104A-hGHbp immobilized on an immunosorb plate (at a concentration of 50 μg/ml) by incubating overnight at 4°C in phosphate-buffered saline solution. Sorting against W104A-hGHbp was continued for seven rounds before individual phagemids were sequenced (9).
  • 17
    • 0029619255 scopus 로고
    • B. C. Cunningham, D. L. Lowe, B. Li, B. D. Bennett, J. A. Wells, EMBO J. 13, 2508 (1994); B. Li et al., Science 270, 1657 (1995).
    • (1995) Science , vol.270 , pp. 1657
    • Li, B.1
  • 19
    • 0023270946 scopus 로고
    • A1-hGH and the single-site revertants were constructed by mutagenizing pB0720, a G120R-hGH expression plasmid (7). These were expressed as soluble proteins [C. N. Chang, M. Rey, B. Bochner, H. Heyneker, G. Gray, Gene 55, 189 (1987)] in the strain 34B8, a nonsuppressor strain of Escherichia coli. Each mutant was isolated from a 50-ml culture in 250-ml shake flasks by pelleting the cells and freezing them at -20°C overnight. Cells were thawed and osmotically shocked in 10 mM tris (pH 8.0).
    • (1987) Gene , vol.55 , pp. 189
    • Chang, C.N.1    Rey, M.2    Bochner, B.3    Heyneker, H.4    Gray, G.5
  • 20
    • 0025944815 scopus 로고
    • The concentration of hormone was determined by densitometry of Coomassie blue-stained gels (4). Initial characterization of binding was measured on a BIA-core instrument by coupling W104A-hGHbp through lysine residues to 2500 resonance units [(4); B. Johnsson, S. Lofas, G. Lindquist, Anal. Biochem. 198, 268 (1991)]. Radioimmunoassay was performed for binding of hGH variants to hGHbp or W104A-hGHbp as described
    • (1991) Anal. Biochem. , vol.198 , pp. 268
    • Johnsson, B.1    Lofas, S.2    Lindquist, G.3
  • 21
    • 0024278671 scopus 로고
    • d measurements ranged from ±30 to ±50% of the value indicated in the text and were higher near the detection limits of the assay (∼1 μM).
    • (1988) J. Biol. Chem. , vol.263 , pp. 7862
    • Spencer, S.A.1
  • 22
    • 1842414407 scopus 로고    scopus 로고
    • note
    • We purified A1-hGH and W104A-hGHbp from an E. coli fermentation culture using ion exchange and hydrophobic interaction columns. The two proteins were combined and purified as a complex on a gel filtration column. Crystals were induced by streak seeding with crystals of the F25A, Y42A, Q46A-hGH:hGHbp complex (75) (∼5 mg/ml) in 50 mM bis-tris (pH 6.5) and 17 to 20% saturated ammonium sulfate. Crystals were frozen in liquid nitrogen for data collection.
  • 23
    • 0026567907 scopus 로고
    • A. E. Ericksson et al., Science 255, 178 (1992); A. E. Ericksson, W. A. Baase, B. W. Matthews, J. Mol. Biol. 229, 747 (1993).
    • (1992) Science , vol.255 , pp. 178
    • Ericksson, A.E.1
  • 27
    • 0029067876 scopus 로고
    • W. Somers, M. Ultsch, A. M. De Vos, A. A. Kossiakoff, Nature 372, 478 (1994); M. R. Walter et al., ibid. 376, 230 (1995); O. Livnah et al., Science 273, 464 (1996).
    • (1995) Nature , vol.376 , pp. 230
    • Walter, M.R.1
  • 28
    • 0029798402 scopus 로고    scopus 로고
    • W. Somers, M. Ultsch, A. M. De Vos, A. A. Kossiakoff, Nature 372, 478 (1994); M. R. Walter et al., ibid. 376, 230 (1995); O. Livnah et al., Science 273, 464 (1996).
    • (1996) Science , vol.273 , pp. 464
    • Livnah, O.1
  • 32
    • 0002452464 scopus 로고
    • L. Sawyer, N. Isaacs, S. Bailey, Eds. SERC Daresbury Laboratory, Warrington, UK
    • Z. Otwinowski, in Data Collection and Processing, L. Sawyer, N. Isaacs, S. Bailey, Eds. (SERC Daresbury Laboratory, Warrington, UK, 1993), p. 56-62.
    • (1993) Data Collection and Processing , pp. 56-62
    • Otwinowski, Z.1
  • 35
    • 1842337602 scopus 로고    scopus 로고
    • note
    • S.A. was supported in part by a NIH Biotechnology Training Grant. We thank the oligonucleotide synthesis and fermentation groups at Genentech; T. Clackson for providing samples of hGHbp and W104A-hGHbp; M. Randal and T. Kossiakoff for help with x-ray data collection; and W. DeLano for help with intermolecular distance calculations. X-ray coordinates have been deposited in the Protein Data Bank with access number 1AXI.


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.