Binding of 5,5'-bis[8-(phenylamino)-1-naphthalenesulfonate] by the regulatory subunits of adenosine cyclic 3',5'-phosphate dependent protein kinase

Bohnert, Janice L.; Malencik, Dean A.; Anderson, Sonia R.; Teller, David C.; Fischer, Edmond H.

doi:10.1021/bi00265a029

Cited by 11 publications

(5 citation statements)

References 19 publications

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“…IDyN ucleotides Inhibit the Bis-ANS-induced Conformational Change in DnaK. Previous studies have suggested that ANS and bis-ANS dyes bind to nucleotide binding sites and could be displaced by nucleotides (Rosen & Weber, 1969;Takashi etal., 1977;Secnik et al, 1990Secnik et al, ,1992Yoo etal., 1990;Griess et al, 1991;Lee et al, 1991;Horowitz et al, 1984;Bohnert et al, 1982;Anderson, 1971). In contrast, our observations suggest that bis-ANS induces a conformational change in DnaK and that nucleotides can reverse the effect of bis-ANS by shifting the equilibrium between native and compact intermediate rather than displacing the bound bis-ANS.…”

Section: Discussioncontrasting

confidence: 54%

“…In addition, our finding that nucleotides can inhibit such bis-ANS-induced perturbation by stabilizing the native state suggests that care should be given to interpreting observations that the addition of nucleotide results in a change in bis-ANS fluorescence of other proteins (Rosen & Weber, 1969;Takashi etal., 1977;Secniketal., 1990;Lee etal., 1991;Bohnert et al, 1982;Anderson, 1971;Dryden et al, 1992; Wiksell & Larsson-Raznikiewicz, 1982). The possibility of a dye-induced conformational change thus offers an alternative interpretation to the hypothesis that bis-ANS is displaced by nucleotide in the same site (Takashi et al, 1977;Yoo et al, 1990;Secnik et al, 1990;Lawson & York, 1987), since for most of the cases the nucleotide did not completely reverse the ANS or bis-ANS fluorescence enhancement, as shown in Table 1.…”

Section: Discussionmentioning

confidence: 94%

“…0006-2960/94/0433-7536S04. 50/0 etal., 1977;Secniketal., 1990Secniketal., ,1992Lee etal., 1991;Horowitz et al, 1984;Bohnert et al, 1982;Anderson, 1971). This assumption is based on the observation that on addition of the nucleotide the fluorescence enhancement of the dye is reduced.…”

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confidence: 99%

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Protein Conformational Changes Induced by 1,1'-Bis(4-anilino-5-naphthalenesulfonic acid): Preferential Binding to the Molten Globule of DnaK

Shi¹,

Palleros²,

Fink³

1994

Biochemistry

131

102

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1,1'-Bis(4-anilino-5-naphthalenesulfonic acid) (bis-ANS), a hydrophobic fluorescent molecular probe which has been shown to bind to compact intermediate states of proteins (molten globules) and also to many nucleotide binding sites, induces a conformational change in DnaK by preferentially binding to its partially folded intermediate state (I) and thus shifting the equilibrium from favoring the native state (N) to favoring the I state. The conformational change was detected by CD, fluorescence emission, size exclusion chromatography, and small-angle X-ray scattering. The presence of bis-ANS significantly decreases the midpoint, Tm, of the initial transition (N-->I) in the thermal unfolding of DnaK, resulting in the apparent destabilization of the native state of DnaK. There is a linear correlation between the apparent free energy (reflected by Tm) of this transition and the concentration of bis-ANS. Bis-ANS does not affect the midpoint of the transition for DnaK from the intermediate to the unfolded state (U). An additional small transition from I to I*, a more expanded intermediate state, was observed, suggesting that the thermal denaturation of DnaK proceeds via a four-state (N-->I-->I*-->U) unfolding process. The addition of nucleotides, ADP or ATP, to the DnaK-bis-ANS complex causes a decrease in bis-ANS fluorescence emission due to the release of bound bis-ANS from the intermediate state of DnaK. This is due to preferential binding of the nucleotide to the native state of DnaK, resulting in a shift in the equilibrium from the intermediate toward the native state rather than the direct displacement of bis-ANS bound in the nucleotide binding site. Denaturation of DnaK induced by bis-ANS can be minimized by working at a temperature much lower than the Tm of the protein, at low dye concentration, and in the presence of nucleotide. Under these conditions, bis-ANS binds to the native state of DnaK.

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Section: Discussioncontrasting

confidence: 54%

Section: Discussionmentioning

confidence: 94%

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Protein Conformational Changes Induced by 1,1'-Bis(4-anilino-5-naphthalenesulfonic acid): Preferential Binding to the Molten Globule of DnaK

Shi¹,

Palleros²,

Fink³

1994

Biochemistry

131

102

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“…* Address correspondence to this author. 0006-2960/86/0425-0739$01.50/0 Bis-ANS has been shown to bind at more than one site in several proteins (Daniel & Weber, 1966;Anderson, 1971;Takashi et al, 1977;Bohnert et al, 1982). The experiments described herein demonstrate the presence of more than one site for the binding of Bis-ANS to tubulin and that additional binding sites appear as a function of time and temperature.…”

mentioning

confidence: 66%

Bis(8-anilinonaphthalene-1-sulfonate) as a probe for tubulin decay

Prasad

Ludueña²,

Horowitz

1986

Biochemistry

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The fluorescent apolar probe bis(8-anilinonaphthalene-1-sulfonate) (Bis-ANS) has been used to detect structural correlates of the well-known but poorly understood decay of tubulin function, by which tubulin loses its ability to polymerize and bind drugs in a complex, time-dependent way. The present results indicate that the decay of tubulin is accompanied by the appearance of hydrophobic areas, which bind a total of six Bis-ANS molecules with a dissociation constant of 19 microM. This binding seems to be a result of localized structural changes that are taking place in the tubulin molecule and can be used as a probe for these changes. In particular, circular dichroism measurements revealed no significant changes in the average secondary structure of the protein during the time required for complete binding of the Bis-ANS molecules. Preincubation of tubulin with the antimitotic drugs colchicine, podophyllotoxin, and vinblastine slows the rate of appearance of the hydrophobic region. Vinblastine has the maximal effect followed by colchicine and podophyllotoxin. In contrast, preincubation with maytansine has no effect. In addition, lowering the temperature decreases the rate of appearance of this region. These results correlate with the effect of drugs on the alkylation of tubulin sulfhydryl groups by iodoacetamide [Luduena, R.F., & Roach, M.C. (1981) Biochemistry 20, 4444-4450] and with the ability of inhibitors of microtubule assembly to permit the polymerization of tubulin into nonmicrotubule structures.

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“…However, neither bis(ANS) nor the parent molecule 8-anilino-1-naphthalene-sulfonate affects nucleotide binding by the isolated catalytic subunit (29). 1,N 6 -Ethenoadenosine cyclic 3Ј,5Ј-phosphate and lin-benzoadenosine 5Ј-phosphate are fluorescent nucleotide derivatives that have been applied to competitive displacement experiments with cAMP and the regulatory subunits (30,31) and with ATP and the isolated catalytic subunit (32), respectively.…”

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confidence: 99%

Binding of 9-Anthroylcholine Monitors the Interactions of Adenosine Cyclic 3′,5′-Phosphate-dependent Protein Kinase with MgATP, Substrates, and Regulatory Subunits

Malencik

Anderson

1998

Journal of Biological Chemistry

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The isolated catalytic subunit of cAMP-dependent protein kinase and smooth muscle myosin light chain kinase undergo interactions with the fluorescent dye 9-anthroylcholine (9AC) that are responsive to the two enzymes' associations with substrates and effectors. Additionally, the binding of 9AC is highly sensitive to subtle structural or functional differences among closely related protein kinases. Skeletal muscle myosin light chain kinase and the catalytically active chymotryptic fragment of the ␥-subunit of phosphorylase kinase do not associate with 9AC. Phosphorylation and dephosphorylation are ubiquitously occurring protein modifications that function in the control of diverse cellular processes. Protein phosphorylation was discovered in conjunction with studies on the enzymology of glycogen degradation. Phosphorylase kinase, which catalyzes the phosphorylation of glycogen phosphorylase (1-3), and cAMP-dependent protein kinase, whose many functions include the phosphorylation and activation of phosphorylase kinase (4 -7), were the first protein kinases to be identified. The two enzymes belong to a large family of eukaryotic protein kinases which share a common "kinase catalytic core" but exhibit varying regulatory mechanisms and specificities (8 -10). Regulation of the individual enzymes often involves specific association with another protein. The holoenzyme of cAMP-dependent protein kinase contains two catalytic subunits and two regulatory subunits, each of which binds two molecules of cAMP. Association with cAMP causes the enzyme tetramer to dissociate into two catalytically active 40.8-kDa subunits (C) and a dimeric regulatory subunit-cAMP complex (5). This reversible reaction is a major component in cAMP-mediated signal transduction. There are two principal types of regulatory subunit, R I (43 kDa) and R II (45 kDa), which differ in both their tissue distributions and functional properties (11)(12)(13)(14).The x-ray crystallographic analyses that were conducted with the isolated catalytic subunit of cAMP-dependent protein kinase demonstrated the binding of a peptide substrate analogue at a site near the entrance to a bilobal cleft, which contains the ATP-binding region. One of the two lobes functions principally in nucleotide binding while the other larger lobe participates in peptide binding and catalysis (15)(16)(17). Crystal studies with an overexpressed truncated form of the catalytically active ␥ subunit of phosphorylase kinase, which possesses 33% sequence identity with the catalytic core (residues 42-297) of cAMP-dependent protein kinase, revealed a similar three-dimensional structure (18). The "catalytic cores" of still other protein kinases have been modeled from the coordinates established by Knighton et al. (15). These enzymes include smooth muscle myosin light chain kinase, which exhibits 30% sequence identity with the aligned 261-residue sequence of cAMP-dependent protein kinase; skeletal muscle myosin light chain kinase, which displays 46% identity with the aligned sequence of the smooth mu...

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Binding of 5,5'-bis[8-(phenylamino)-1-naphthalenesulfonate] by the regulatory subunits of adenosine cyclic 3',5'-phosphate dependent protein kinase

Cited by 11 publications

References 19 publications

Protein Conformational Changes Induced by 1,1'-Bis(4-anilino-5-naphthalenesulfonic acid): Preferential Binding to the Molten Globule of DnaK

Protein Conformational Changes Induced by 1,1'-Bis(4-anilino-5-naphthalenesulfonic acid): Preferential Binding to the Molten Globule of DnaK

Bis(8-anilinonaphthalene-1-sulfonate) as a probe for tubulin decay

Binding of 9-Anthroylcholine Monitors the Interactions of Adenosine Cyclic 3′,5′-Phosphate-dependent Protein Kinase with MgATP, Substrates, and Regulatory Subunits

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