Inhibition of Escherichia coli RNA polymerase by bis(1-anilino-8-naphthalenesulfonate)

Wu, Fan; Wu, Cheng‐Wen

doi:10.1021/bi00594a020

Cited by 15 publications

(8 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The data presented in Figures 5 and 6 suggest that bis-ANS and nucleic acid bind to the same region of the NC protein. This is supported by reports that bis-ANS has a particular affinity for proteins that have nucleotide binding sites (Worah et al, 1978;Wu & Wu, 1978;Prasad et al, 1986;Lin et al, 1983;Holler et al, 1971). The tight binding between the NC protein and poly (A), as measured by the displacement of bis-ANS, is consistent with previous observations, as is the site size of 6 (Jentoft et al, 1988).…”

Section: Discussionsupporting

confidence: 89%

See 1 more Smart Citation

Interactions at the nucleic acid binding site of the avian retroviral nucleocapsid protein: studies utilizing the fluorescent probe 4,4'-bis(phenylamino)(1,1'-binaphthalene)-5,5'-disulfonic acid

Secnik¹,

Wang²,

Chang³

et al. 1990

Biochemistry

View full text Add to dashboard Cite

The structural and functional properties of the nucleocapsid (NC) protein of the avian myeloblastosis virus were examined by steady-state fluorescence and fluorescence anisotropy measurements of the complex between the NC and the extrinsic fluorophore 4,4'-bis(phenylamino)(1,1'-binaphthalene)-5,5'-disulfonic acid (bis-ANS). The intrinsic fluorescence of bis-ANS is enhanced many fold upon forming a complex with the NC. Between 2 and 10 molecules of bis-ANS bind strongly to the NC, with an overall Kd of less than 10(-6) M. The emission of bis-ANS in the complex can also be induced by excitation at 298 nm, indicating that energy is transferred from Trp 80, the sole tryptophan in the NC protein, to bis-ANS. The energy transferred between the Trp 80 and bis-ANS was analyzed to yield a calculated distance of separation between these fluorophores of 28 +/- 3 A; thus, Trp 80 is well removed from the nearest bound bis-ANS. The fluorescence emission of bis-ANS in the NC.bis-ANS complex is efficiently quenched by added salts and by poly(A), suggesting that salt (presumably anions), nucleic acid, and bis-ANS bind to the same, positively charged region on the NC protein. A site size of six nucleotides was determined for nucleic acid binding to the NC protein, with an estimated Kd of less than 10(-6) M. Salt (anion) binding is strong, but nonspecific, with a Kapp of 4 mM, raising the possibility that anion binding to the NC protein might regulate the interaction of the NC with viral RNA inside the host cell.

show abstract

Section: Discussionsupporting

confidence: 89%

“…Multiple binding sites for bis-ANS have been reported for other proteins such as RNA polymerase figure 6: Plot of the quenching of the bis-ANS fluorescence in the NC-bis-ANS complex by poly(A) (representative data shown in Figure 5). (Wu & Wu, 1978), E. coli lac repressor (Worah et al, 1978), and tubulin (Prasad et al, 1986).…”

Section: Discussionmentioning

confidence: 99%

Interactions at the nucleic acid binding site of the avian retroviral nucleocapsid protein: studies utilizing the fluorescent probe 4,4'-bis(phenylamino)(1,1'-binaphthalene)-5,5'-disulfonic acid

Secnik¹,

Wang²,

Chang³

et al. 1990

Biochemistry

View full text Add to dashboard Cite

show abstract

“…In addition to being a hydrophobic probe, other reactions of ANS have been reported, e.g. its stimulatory effect on the catalytic activity of cytochrome P-450 [39], its inhibitory effect on RNA polymerase [40] and its denaturing effect on DnaK [41].…”

Section: Discussionmentioning

confidence: 99%

Binding of a burst-phase intermediate formed in the folding of denatured D-glyceraldehyde-3-phosphate dehydrogenase by chaperonin 60 and 8-anilino-1-naphthalenesulphonic acid

Lei

Cai

et al. 1998

Biochemical Journal

View full text Add to dashboard Cite

Upon dilution, D-glyceraldehyde-3-phosphate dehydrogenase (GADPH) that has been fully inactivated, but only partially unfolded, in dilute guanidine hydrochloride (GuHCl) recovers activity completely. The fully unfolded enzyme, however, is re-activated only to a limited extent after dilution, and refolds rapidly in a burst phase to a partially folded intermediate characterized by increases in both the emission intensity of intrinsic fluorescence and binding to 8-anilino-1-naphthalenesulphonic acid (ANS). This intermediate aggregates with a time lag of a few minutes, and the aggregation can be suppressed completely by chaperonin 60 (GroEL). Stoichiometric analysis of the suppression of GAPDH re-activation by GroEL suggests that the tetradecameric GroEL binds to a dimeric GAPDH folding intermediate. This intermediate can be re-activated by ATP or ATP/chaperonin 10 (GroES) to an extent considerably greater than that obtained on spontaneous re-activation of the fully denatured enzyme upon dilution. Probing with a fluorescent derivative of NAD+ shows that this folding intermediate does not have a native conformation at the active site. The similar profiles of the effects of GroEL and ANS on the re-activation of GAPDH denatured by different concentrations of GuHCl suggest that GroEL and ANS recognize and bind to the same folding intermediate, which is similar to the relatively stable, partially unfolded, state of the enzyme denatured in 0.5-1.0 MGuHCl. However, the complexes of the intermediate with GroEL or ANS appear to be different, in that GroEL, but not ANS, suppresses aggregation and assists folding in the presence of ATP.

show abstract

“…The structure of bis-Ans has been determined (Farris et al, 1978) and it has been shown to be a particularly useful probe of nucleotide binding sites on proteins. Those proteins examined using bis-Ans include glutamate dehydrogenase (Anderson, 1971), the myosin ATPase (Takashi et al, 1977), and RNA polymerase (Wu & Wu, 1978). This paper is the first reported use of bis-Tns as a probe of protein conformation.…”

Section: Discussionmentioning

confidence: 99%

“…Although it is too soon to make a general statement, it appears that, upon binding, these large aromatic probes can change the protein conformation in such a way as to unfold or denature the protein, opening the way to further dye binding. Dye binding studies with pyruvate oxidase, glutamate dehydrogenase (Anderson, 1971), and RNA polymerase (Wu & Wu, 1978) indicate large quantities of the probe binding to these proteins. Despite the fact that the binding of bis-Tns to pyruvate oxidase is not to a well-defined site, such as the cofactor binding site, the interaction between the probe and the enzyme has been useful in exploring this complicated membrane enzyme.…”

Section: Discussionmentioning

confidence: 99%

Studies on the interaction between Escherichia coli pyruvate oxidase and a detergent activator by utilization of the fluorescence probe bis(8-p-toluidino-1-naphthalene)sulfonate

O'Brien

Gennis²

1979

Biochemistry

View full text Add to dashboard Cite

Pyruvate oxidase is a peripheral membrane flavoenzyme isolated from Escherichia coli. The in vitro specific activity of the pure enzyme is enhanced 25-fold in the presence of certain lipids and detergents. In addition, the affinity of the protein for both phospholipids and detergents is significantly enhanced in the presence of the enzyme substrate and cofactor, pyruvate and thiamin pyrophosphate (Mg2'). In this paper a novel fluorescent probe is used to examine the protein conformational changes concomitant with substrate reduction of the flavin, activation of the oxidase, and binding of the detergent activator, dodecyl sulfate. In the presence of dodecyl sulfate, the probe bis(8-p-toluidino-1 -naphthalenesulfonate) (bis-Tns) binds to pyruvate oxidase with a resultant large increase in probe fluorescence at dye concentrations below 1 pM. No binding at low dye concentration is observed in the absence of the activator. The enzyme binds about 16 molecules of bis-Tns, in what appear to be hydrophobic binding sites. The substrate-reduced flavoprotein binds to bis-Tns not only

show abstract

Inhibition of Escherichia coli RNA polymerase by bis(1-anilino-8-naphthalenesulfonate)

Cited by 15 publications

References 23 publications

Interactions at the nucleic acid binding site of the avian retroviral nucleocapsid protein: studies utilizing the fluorescent probe 4,4'-bis(phenylamino)(1,1'-binaphthalene)-5,5'-disulfonic acid

Interactions at the nucleic acid binding site of the avian retroviral nucleocapsid protein: studies utilizing the fluorescent probe 4,4'-bis(phenylamino)(1,1'-binaphthalene)-5,5'-disulfonic acid

Binding of a burst-phase intermediate formed in the folding of denatured D-glyceraldehyde-3-phosphate dehydrogenase by chaperonin 60 and 8-anilino-1-naphthalenesulphonic acid

Studies on the interaction between Escherichia coli pyruvate oxidase and a detergent activator by utilization of the fluorescence probe bis(8-p-toluidino-1-naphthalene)sulfonate

Contact Info

Product

Resources

About