Reaction of the adenine nucleotide analogue 5'-p-fluorosulfonylbenzoyl adenosine at distinct tyrosine and cysteine residues of rabbit muscle pyruvate kinase.

Annamalai, Anjanayaki E.; Colman, Robertal F.

doi:10.1016/s0021-9258(19)68616-3

Cited by 62 publications

(20 citation statements)

References 40 publications

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“…Sulfo-7V-succinimidyl esters in aqueous solutions react rapidly and in high yield with protein amino groups to form stable products (Anjaneyulu & Staros, 1987). The fluorosulfonyl group which comprises the second moiety of this cross-linker, however, is relatively less reactive in aqueous solutions (Limbird et al, 1983) 1982), tyrosyl (Esch & Allison, 1978;Likos et al, 1980;Annamalai & Colman, 1981), and, by analogy to the chlorosulfonyl group, histidinyl and activated seryl (Paulos & Price, 1974) residues when bound at a site containing such a nucleophilic group.…”

Section: Resultsmentioning

confidence: 99%

High-yield covalent attachment of epidermal growth factor to its receptor by kinetically controlled, stepwise affinity cross-linking

et al. 1992

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We report here the use of a stepwise affinity cross-linking technique in the specific covalent attachment of epidermal growth factor (EGF) to its receptor. A heterobifunctional cross-linking reagent, sulfo-N-succinimidyl 4-(fluorosulfonyl)benzoate (SSFSB), which contains a rapidly reacting sulfo-N-succinimidyl active ester and a much more slowly reacting aromatic fluorosulfonyl moiety, was synthesized and characterized. Murine EGF (mEGF) was modified by the cross-linker to yield as the major product a derivative of mEGF having the (fluorosulfonyl)benzoyl moiety attached covalently at the amino terminus. SSFSB-modified, 125I-labeled mEGF was separated from unreacted SSFSB by size-exclusion chromatography and applied to shed membrane vesicles from A431 human carcinoma cells. Binding of derivatized 125I-mEGF to vesicles led to high yields (greater than 60%) of covalent linkage of 125I-mEGF to the EGF receptor, as determined by measurement of the fraction of specifically bound radiolabel which comigrated with the EGF receptor in NaDodSO4-polyacrylamide gels. The specificity of affinity cross-linking was evident in the negligible degree of labeling of species other than the EGF receptor and in the retention of EGF-stimulated receptor kinase activity after cross-linking.

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Section: Resultsmentioning

confidence: 99%

High-yield covalent attachment of epidermal growth factor to its receptor by kinetically controlled, stepwise affinity cross-linking

et al. 1992

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“…To our knowledge, these data constitute the first report of the specific induction of a cystine disulfide by a photolyzed aryl azide. It is of interest to note that the inactivation of pyruvate kinase by 5'-[/?-(fluorosulfonyl)benzoyl]adenosine occurs through a similar mechanism, in which the formation of an intramolecular cystine disulfide causes displacement of label (Annamalai & Colman, 1981). A number of nucleotide-binding proteins are known to have reactive or critical cysteine residues within or near their active sites.…”

Section: Discussionmentioning

confidence: 99%

Formation of an intramolecular cystine disulfide during the reaction of 8-azidoguanosine 5'-triphosphate with cytosolic phosphoenolpyruvate carboxykinase (GTP) causes inactivation without photolabeling

Lewis

Haley²,

Carlson³

1989

Biochemistry

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Phosphoenolpyruvate carboxykinase (GTP) (PEPCK) specifically utilizes a guanosine or inosine nucleotide as a substrate, yet it does not share extended sequence homology with other GTP-binding proteins, and the molecular basis for its nucleotide specificity is not understood. In an effort to locate the enzyme's nucleotide-binding site, we have studied the interaction of cytosolic PEPCK from rat liver with the photoprobe 8-azidoGTP, which fulfills the criteria of a specific photoaffinity label for PEPCK. The photoprobe binds reversibly to the enzyme prior to modification and at low concentrations causes greater than 60% inactivation (Ki = 1.2 microM). GTP provides nearly complete protection against inactivation by 8-azidoGTP, whereas phosphoenolpyruvate and metal ions provide partial protection. In addition, the photoprobe is a substrate for the enzyme and has a Km similar to that for GTP. However, the extent of covalent modification by [32P]8-azidoGTP as measured by three independent techniques is significantly lower than the extent of enzyme inactivation. Further investigation of this anomaly has revealed that the loss in enzymatic activity is caused by modification of a critical cysteine residue in a reaction that does not terminate with covalent attachment of the photolabel. Quantitation of the total free thiols of modified PEPCK shows that 2 mol of cysteine is lost per mole of inactivated enzyme. These results indicate that the photoinactivation of PEPCK by 8-azidoGTP is caused by the formation of an intramolecular cystine disulfide bridge, thus providing evidence for the existence of a pair of proximal cysteine residues within the GTP-binding site. The interaction of cysteine residues with the reactive photogenerated derivatives of 8-azidopurines is discussed.

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“…For example, it was postulated that an ATP-site cysteine in rabbit muscle pyruvate kinase reacts with FSBA to form a thiosulfonate intermediate that is subsequently quenched by an additional neighbouring cysteine, thus forming a partially active disulde enzyme. 93 Similarly, isocitrate dehydrogenase reacted with FSBA to form a disulde bond that inhibited ATPase activity. 94 PMSF was found to react with the N-terminal proline amine group in macrophage migration inhibitory factor (MIF) unexpectedly (Fig.…”

Section: Other Residuesmentioning

confidence: 99%

Sulfonyl fluorides as privileged warheads in chemical biology

2015

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Reaction of the adenine nucleotide analogue 5'-p-fluorosulfonylbenzoyl adenosine at distinct tyrosine and cysteine residues of rabbit muscle pyruvate kinase.

Cited by 62 publications

References 40 publications

High-yield covalent attachment of epidermal growth factor to its receptor by kinetically controlled, stepwise affinity cross-linking

High-yield covalent attachment of epidermal growth factor to its receptor by kinetically controlled, stepwise affinity cross-linking

Formation of an intramolecular cystine disulfide during the reaction of 8-azidoguanosine 5'-triphosphate with cytosolic phosphoenolpyruvate carboxykinase (GTP) causes inactivation without photolabeling

Sulfonyl fluorides as privileged warheads in chemical biology

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