-
1
-
-
0025123333
-
-
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
-
-
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
-
-
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
-
-
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
-
-
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
-
-
-
9
-
-
0025765004
-
-
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).
-
(1991)
Proc. Natl. Acad. Sci. U.S.A.
, vol.88
, pp. 4498
-
-
Bass, S.H.1
Mulkerrin, M.G.2
Wells, J.A.3
-
10
-
-
0028916599
-
-
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
-
-
note
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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.
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12
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0025888264
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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
-
-
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
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14
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1842298302
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note
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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).
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16
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0028239263
-
-
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).
-
(1994)
EMBO J.
, vol.13
, pp. 2508
-
-
Cunningham, B.C.1
Lowe, D.L.2
Li, B.3
Bennett, B.D.4
Wells, J.A.5
-
17
-
-
0029619255
-
-
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
-
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0023270946
-
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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
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20
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0025944815
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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
-
-
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
-
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-
-
1842414407
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note
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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.
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-
23
-
-
0026567907
-
-
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
-
24
-
-
0027394606
-
-
A. E. Ericksson et al., Science 255, 178 (1992); A. E. Ericksson, W. A. Baase, B. W. Matthews, J. Mol. Biol. 229, 747 (1993).
-
(1993)
J. Mol. Biol.
, vol.229
, pp. 747
-
-
Ericksson, A.E.1
Baase, W.A.2
Matthews, B.W.3
-
25
-
-
0029794909
-
-
K. H. Pearce, M. H. Ultsch, R. F. Kelley, A. M. De Vos, J. A. Wells, Biochemistry 35, 10300 (1996).
-
(1996)
Biochemistry
, vol.35
, pp. 10300
-
-
Pearce, K.H.1
Ultsch, M.H.2
Kelley, R.F.3
De Vos, A.M.4
Wells, J.A.5
-
26
-
-
0028032203
-
-
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).
-
(1994)
Nature
, vol.372
, pp. 478
-
-
Somers, W.1
Ultsch, M.2
De Vos, A.M.3
Kossiakoff, A.A.4
-
27
-
-
0029067876
-
-
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
-
-
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
-
31
-
-
0027467033
-
-
L. J. Keefe, J. Sondek, D. Shortle, E. E. Lattman, Proc. Natl. Acad. Sci. U.S.A. 90, 3275 (1993).
-
(1993)
Proc. Natl. Acad. Sci. U.S.A.
, vol.90
, pp. 3275
-
-
Keefe, L.J.1
Sondek, J.2
Shortle, D.3
Lattman, E.E.4
-
32
-
-
0002452464
-
-
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
-
-
note
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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.
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