An Environmentally-Sensitive Reagent with Selectivity for the Tryptophan Residue in Proteins

Koshland, Daniel E.; Karkhanis, Yashwant D.; Latham, H. George

doi:10.1021/ja01061a045

Cited by 172 publications

(51 citation statements)

References 2 publications

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“…Along this line, selective chemical modifications of strategic amino acids in the ADP/ATP carrier have already provided interesting data [4,5,8]. The penetrant reagents used so tar and their main targets include NEM (SH groups) [91, HNB (tryptophanyl residues) [10], phenylglyoxal and butanedione (arginyl residues [ 11,12]). Inactivation of the binding properties of the ADP/ATP carrier by UV light was possibly due to modification of tryptophanyl residues [4].…”

Section: Introductionmentioning

confidence: 99%

Atractyloside and bongkrekic acid sites in the mitochondrial ADP/ATP carrier protein

1981

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Section: Introductionmentioning

confidence: 99%

Atractyloside and bongkrekic acid sites in the mitochondrial ADP/ATP carrier protein

1981

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“…The effect of bound polypeptide on ATP-induced structural rearrangements in GroEL was then assessed using the intrinsic fluorescence of Trp-485-GroEL. However, in this case, to avoid contamination of the fluorescence signal by the presence of the four native tryptophan residues in ␣-lactalbumin, the CM-LA was treated with Koshland's reagent (HNBB), which forms a non-fluorescent adduct with the indole group so that substrate fluorescence does not interfere with the indole signal from Trp-485 in GroEL (37).…”

mentioning

confidence: 99%

Elucidation of Steps in the Capture of a Protein Substrate for Efficient Encapsulation by GroE

Cliff¹,

Limpkin²,

Cameron

et al. 2006

Journal of Biological Chemistry

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We have identified five structural rearrangements in GroEL induced by the ordered binding of ATP and GroES. The first discernable rearrangement (designated T 3 R 1 ) is a rapid, cooperative transition that appears not to be functionally communicated to the apical domain. In the second (R 1 3 R 2 ) step, a state is formed that binds GroES weakly in a rapid, diffusion-limited process. However, a second optical signal, carried by a protein substrate bound to GroEL, responds neither to formation of the R 2 state nor to the binding of GroES. This result strongly implies that the substrate protein remains bound to the inner walls of the initially formed GroEL⅐GroES cavity, and is not yet displaced from its sites of interaction with GroEL. In the next rearrangement (R 2 ⅐GroES 3 R 3 ⅐GroES) the strength of interaction between GroEL and GroES is greatly enhanced, and there is a large and coincident loss of fluorescence-signal intensity in the labeled protein substrate, indicating that there is either a displacement from its binding sites on GroEL or at least a significant change of environment. These results are consistent with a mechanism in which the shift in orientation of GroEL apical domains between that seen in the apo-protein and stable GroEL⅐GroES complexes is highly ordered, and transient conformational intermediates permit the association of GroES before the displacement of bound polypeptide. This ensures efficient encapsulation of the polypeptide within the GroEL central cavity underneath GroES.

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“…Alkylation of tryptophan residues with nitrophenols. The method of Koshland et al (1964) was adapted. -2-Hydroxy-5-nitrobenzyl bromide (HNBBr) at concentrations ranging from 0.33 to 18 mM was added as a solid or from a 3 mM stock solution (in the same buffer) to a 1-2 pM solution of toxin in 0.1 M acetate/acetic acid, pH 3.0/50/0 (v/v) methanol.…”

Section: Chemical Modifications Of Cryla(c) Toxinmentioning

confidence: 99%

Chemical modification of Bacillus thuringiensis activated -endotoxin and its effect on toxicity and binding to Manduca sexta midgut membranes

Cumming

1994

Microbiology

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The effect of chemical modification of aromatic and basic residues of the Bacillus thuringiensis insecticidal CrylA(c) toxin on toxicity and receptor binding was studied. Modification of four or more of the 43 arginine residues resulted in abolition of toxicity towards Manduca sexfa and Pieris brassicae in vivo and a Choristoneura fumiferana cell line in witro. Modification of seven or more of the 27 tyrosine residues resulted in reduction of toxicity. Upon modification of most of the 10 tryptophan residues, toxicity was reduced. Modification of histidine residues had no effect on toxicity. A quantitative binding assay was developed and optimized to compare the binding of derivatives with native toxin t o M. sexta brush border membrane vesicles. The reduction or abolition of toxicity observed upon selective modification of tyrosine or arginine residues was reflected in the binding abilities of the derivatives. However a non-toxic derivative, modified at tryptophan residues, retained its ability to bind vesicles. These results suggest that tyrosine and arginine residues are involved in the binding interaction of toxin with receptor but tryptophan residues are involved in some subsequent step.

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An Environmentally-Sensitive Reagent with Selectivity for the Tryptophan Residue in Proteins

Cited by 172 publications

References 2 publications

Atractyloside and bongkrekic acid sites in the mitochondrial ADP/ATP carrier protein

Atractyloside and bongkrekic acid sites in the mitochondrial ADP/ATP carrier protein

Elucidation of Steps in the Capture of a Protein Substrate for Efficient Encapsulation by GroE

Chemical modification of Bacillus thuringiensis activated -endotoxin and its effect on toxicity and binding to Manduca sexta midgut membranes

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