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2
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0344992646
-
-
note
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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; Y, Tyr.
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2-terminal residues (four vector-derived and TfR 117 to 120) as follows. The TfR (4 mg) was concentrated by vacuum dialysis against 100 mM KCl, 100 mM NaCl, 5 mM potassium phosphate (pH 6.7) (buffer A) to a final concentration of 1 to 2 mg/ml. TfR was removed and placed on ice, and 20 ml of trypsin (10 mg/ml) was added. After 1 hour, TfR was passed over a benzamidine Sepharose column, and phenylmethysulfonyl fluoride was added (up to 1 mM) to inhibit residual trypsin activity. We then passed the TfR over a Superdex 200 size-exclusion column equilibrated with buffer A to remove aggregated protein.
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19
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0344130095
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note
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free for the refined model, using data from 8.0 to 3.2 Å with l/σ(l) > 2.0, are 24.0% and 28.7%, respectively. The root-mean-square deviations in bond lengths and bond angles are 0.007 Å and 1.18°. Figs. 1 and 2 were drawn with RIBBONS (29), and Fig. 4 was drawn with GRASP (30).
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We thank D. Thiel and other staff members at CHESS for assistance with data collection at the F-1 beamline; W. Minor for help in data processing; R. Davis for help with CHO cell expression; P. Aisen, N. Andrews, M. Eck, and M. Wessling-Resnick for discussion; and P. Bjorkman, M. Bennet, and C. Enns for communicating results before publication. C.M.L is an Armenise Fellow of the Harvard Medical School Center for Structural Biology.
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