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Abbreviations used: CD, circular dichroism; DTNB, 5,5'-dithiobis-(2-nitrobenzoic) acid; EDTA, ethylenediaminetetracetic acid; GdnHCl, guanidine hydrochloride; GR, yeast glutathione reductase; GSH, reduced glutathione; GSSG, oxidized glutathione; HEPES, n-(2-hydroxyethyl) piperazine-n'-(2-ethanesulfonic acid); HPLC, high performance liquid chromatography; MG, molten globule
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. Titration of reactive thiol groupsThis was performed by following the increase in 2-nitro-5-thiobenzoic acid (TNB) absorbance at 412 nm, according to Collier (1973)
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. Similarly, only 13 out of 53 hydrophobic interactions are intrasubunit in nature
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. Identification of nanosecond-timescale internal motionThe presence of nanosecond-timescale motion can be directly identified from a plot of the normalized ratio of R^sub 1^ values measured at two magnetic fields (RT^sup n^^sub 1^) versus NOE (22)
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. This reduced affinity allows both G?(GTP) and Gß? to bind to and change the catalytic activity of a variety of intracellular effector proteins
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. This probe becomes fluorescent when it reacts with cysteine side chains (see Molecular Probes Handbook, Eugene, OR)
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- Cooperativity induced by a single mutation at the subunit interface of a dimeric enzyme: glutathione reductase
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