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d for each mutant. The 37 mutants tested, and their relative affinities (%) were: E217R (123%), E227R (109%), K242E (92%), E267R (117%), H271R (123%), T277R (7%), T281R (145%), V284R (105%), D285A (89%), K288A (98%), C294K (94%), E295R (118%), C298A (87%), C298R (141%), E299A (171%), I302A (86%), I302R (99%), K306A (6%), K306E (6%), P384R (164%), A387R (107%), E390R (151%), E393R (146%), L400R (95%), H413R (109%), H416R (153%), M423R (156%), R429A (48%), S437R (170%), L440R (174%), V444R (89%), T448R (234%), E449R (36%), P453E (32%), L454R (26%), L456R (46%), and E457K (71%).
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2642611766
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Single-letter abbreviations for the 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
-
Single-letter abbreviations for the 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.
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2642704054
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note
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3. After the beads were washed three times with 1 ml of binding buffer, the bound proteins were separated with use of 10% SDS-PAGE and visualized by autoradiography. Binding was quantitated by phosphorimaging with ImageQuant (Molecular Dynamics).
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37
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2, as indicated. After cell culture for 24 hours, the LUC or CAT activities were assayed; β-Gal activities were also assayed to correct for differences in transfection efficiencies.
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41
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2642610099
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We thank M. Stallcup and C. Costa for providing the GST-GRIP1, GST-RXRα, and GST-TRβ expression vectors; P. Webb and M. Tsai for providing the pSG5-GRIP1 and pCMX-SRC-1a expression vectors; K. Yamamoto, P. Webb, M. G. Rosenfeld, and T. Scanlan for suggestions; and T. Chen and K. Nakamura for technical assistance. W.F. is a recipient of a postdoctoral fellowship from the National Institutes of Health (DK09516). Supported by NIH grants to J.D.B. (DK51281) and to the Computer Graphics Laboratory at the University of California, San Francisco (P41-RR01081)
-
We thank M. Stallcup and C. Costa for providing the GST-GRIP1, GST-RXRα, and GST-TRβ expression vectors; P. Webb and M. Tsai for providing the pSG5-GRIP1 and pCMX-SRC-1a expression vectors; K. Yamamoto, P. Webb, M. G. Rosenfeld, and T. Scanlan for suggestions; and T. Chen and K. Nakamura for technical assistance. W.F. is a recipient of a postdoctoral fellowship from the National Institutes of Health (DK09516). Supported by NIH grants to J.D.B. (DK51281) and to the Computer Graphics Laboratory at the University of California, San Francisco (P41-RR01081).
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