Reactivity of tyrosine in bovine pancreatic deoxyribonuclease with p-nitrobenzenesulfonyl fluoride.

Liao, Ta–Hsiu; Ting, Ruby; Yeung, Jennifer

doi:10.1016/s0021-9258(19)83825-5

Cited by 43 publications

(7 citation statements)

References 34 publications

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“…Since the inhibition of the placental H+ pump by NBD-CI as well as the reversibility of this inhibition by thiols does not distinguish between the participation of tyrosyl groups and that of thiol groups, we investigated the influence of three additional reagents which are commonly used to modify tyrosyl residues in proteins. These reagents are tetranitromethane (Glazer, 1975; Lundbland & Noyes, 1984), pNBSF (Liao et al, 1982), and Nacetylimidazole (Cohen, 1968). Incubation of cholate-pretreated membrane vesicles with all three reagents led to inactivation of the H+ pump.…”

Section: Resultsmentioning

confidence: 99%

The ATP-binding site of the human placental hydrogen ion pump contains essential tyrosyl residues

Kulanthaivel¹,

Simon²,

Burckhardt³

et al. 1990

Biochemistry

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Transient exposure of human placental brush-border membrane vesicles to cholate reorients the ATP-driven H+ pump, enabling the pump to transport H+ into the vesicles upon addition of ATP to the external medium. H+ uptake can be measured in these vesicles by following the decrease in the absorbance of acridine orange, a delta pH indicator. We investigated the role of tyrosyl residues in the catalytic function of the H+ pump by studying the effects of tyrosyl group specific reagents on ATP-driven H+ uptake in cholate-pretreated membrane vesicles. The reagents tested were 7-chloro-4-nitro-2,1,3-benzoxadiazole (NBD-Cl), N-acetylimidazole, tetranitromethane, and p-nitrobenzenesulfonyl fluoride. Treatment of the membrane vesicles with these reagents resulted in the inhibition of the ATP-driven H+ uptake, and the inhibitory potency was in the following order: NBD-Cl greater than tetranitromethane greater than p-nitrobenzenesulfonyl fluoride greater than N-acetylimidazole. The inhibition of the H+ pump by NBD-Cl was reversible by 2-mercaptoethanol, and the inhibition by N-acetylimidazole was reversible by hydroxylamine. Since these reagents are not absolutely specific for tyrosyl groups and can also react with thiol groups, we studied the interaction of N-acetylimidazole with the H+ pump whose triol groups were masked by reaction with p-(chloromercuri)benzenesulfonate. The SH-masked pump was totally inactive, but the activity could be restored by dithiothreitol. On the contrary, the activity of the SH-masked H+ pump which was subsequently treated with N-acetylimidazole could not be restored by dithiothreitol, suggesting that thiol groups were not involved in the inhibition of the H+ pump by N-acetylimidazole.(ABSTRACT TRUNCATED AT 250 WORDS)

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Section: Resultsmentioning

confidence: 99%

The ATP-binding site of the human placental hydrogen ion pump contains essential tyrosyl residues

Kulanthaivel¹,

Simon²,

Burckhardt³

et al. 1990

Biochemistry

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“…Inhibition Studies with NBSF. The DT-diaphorase (0.65 µ ) was incubated with different concentrations of NBSF/ ethanol solution (0-30 µ ) in the absence of coenzyme or substrate at room temperature, as described previously (Liao et al, 1982). At each time interval, one aliquot (10 µL) was taken for enzyme assay, followed by addition of 10 µL of 8 mM NADH and 10 µL of 8 mM menadione in 1.0 mL of 0.1 M phosphate buffer, pH 7.4.…”

Section: Methodsmentioning

confidence: 99%

Structure-function relationship of NAD(P)H:quinone reductase: characterization of amino-terminal blocking group and essential tyrosine and lysine residues

Haniu¹,

Yuan²,

Chen³

et al. 1988

Biochemistry

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The amino terminal blocked peptide of rat liver NAD(P)H:quinone reductase (DT-diaphorase) was determined by amino acid sequence analysis and by mass spectrometry. The mature protein is composed of 273 amino acids and contains an acetylated amino terminus, which was not identified by previous cDNA analysis. The enzyme was inactivated by p-nitrobenzenesulfonyl fluoride (NBSF) or 2,4,6-trinitrobenzenesulfonate (TNBS) with pseudo-first-order kinetics. These studies suggest that essential tyrosine and lysine may be present in the active site of this enzyme. The NBSF inhibition was protected by 1-naphthol and 1-naphthylamine, but not by NAD+. However, TNBS inhibition was not prevented by the naphthalene derivatives or NAD+. Specific peptides labeled with NBSF or TNBS were isolated by high-performance liquid chromatography and were sequenced. These analyses revealed that the NBSF-labeled tyrosine resides in a predominantly hydrophobic region and TNBS-labeled lysine in a predominantly hydrophilic region.

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“…Apart from the three major reagents introduced above, some other reagents for Tyr footprinting are acid anhydrides and acyl chlorides (target phenolic hydroxyl groups), , cyanuric fluoride (targets phenolic hydroxyl groups), , various diazonium salts (target phenolic C–H bonds), , polyhalogenated quinones (target phenolic hydroxyl groups), p -nitrobenzenesulfonyl fluoride (NBSF, targets phenolic hydroxyl group), , diisopropylphosphorofluoridate (targets phenolic hydroxyl groups), , p -nitrophenyl acetate (targets phenolic hydroxyl groups), and hemin-activated luminol derivatives (target phenolic C–H bonds) . Although this wide variety of reactive reagents demonstrates the opportunities of footprinting a given functional group, most await establishment in MS-based protein HOS elucidation.…”

Section: Targeted-labeling Reagentsmentioning

confidence: 99%

Mass Spectrometry-Based Protein Footprinting for Higher-Order Structure Analysis: Fundamentals and Applications

2020

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Proteins adopt different higher-order structures (HOS) to enable their unique biological functions. Understanding the complexities of protein higher-order structures and dynamics requires integrated approaches, where mass spectrometry (MS) is now positioned to play a key role. One of those approaches is protein footprinting. Although the initial demonstration of footprinting was for the HOS determination of protein/nucleic acid binding, the concept was later adapted to MS-based protein HOS analysis, through which different covalent labeling approaches “mark” the solvent accessible surface area (SASA) of proteins to reflect protein HOS. Hydrogen–deuterium exchange (HDX), where deuterium in D2O replaces hydrogen of the backbone amides, is the most common example of footprinting. Its advantage is that the footprint reflects SASA and hydrogen bonding, whereas one drawback is the labeling is reversible. Another example of footprinting is slow irreversible labeling of functional groups on amino acid side chains by targeted reagents with high specificity, probing structural changes at selected sites. A third footprinting approach is by reactions with fast, irreversible labeling species that are highly reactive and footprint broadly several amino acid residue side chains on the time scale of submilliseconds. All of these covalent labeling approaches combine to constitute a problem-solving toolbox that enables mass spectrometry as a valuable tool for HOS elucidation. As there has been a growing need for MS-based protein footprinting in both academia and industry owing to its high throughput capability, prompt availability, and high spatial resolution, we present a summary of the history, descriptions, principles, mechanisms, and applications of these covalent labeling approaches. Moreover, their applications are highlighted according to the biological questions they can answer. This review is intended as a tutorial for MS-based protein HOS elucidation and as a reference for investigators seeking a MS-based tool to address structural questions in protein science.

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Reactivity of tyrosine in bovine pancreatic deoxyribonuclease with p-nitrobenzenesulfonyl fluoride.

Cited by 43 publications

References 34 publications

The ATP-binding site of the human placental hydrogen ion pump contains essential tyrosyl residues

The ATP-binding site of the human placental hydrogen ion pump contains essential tyrosyl residues

Structure-function relationship of NAD(P)H:quinone reductase: characterization of amino-terminal blocking group and essential tyrosine and lysine residues

Mass Spectrometry-Based Protein Footprinting for Higher-Order Structure Analysis: Fundamentals and Applications

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