Effects of Temperature and Activating Cations on the Fluorescence of Pyruvate Kinase<sup>*</sup>

Ch, Suelter

doi:10.1021/bi00854a008

Cited by 41 publications

(30 citation statements)

References 10 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…, increase the solvent exposure of Trp-157. Compared to the previous study, these results clearly indicate that the majority of changes in tryptophan fluorescence signal from PK induced by the binding of activating cations come from Trp-157 (Kayne and Suelter 1965;Ou et al 2010;Suelter 1967).…”

Section: Interaction With Activating Cationscontrasting

confidence: 50%

“…A variety of techniques have been used to explore changes in the conformation of pyruvate kinase in response to ligand binding (Carminat et al 1971; Kayne and Price 1972;Kayne and Suelter 1965;Cohn 1965, 1966;Oria-Hernandez et al 2005;Ou et al 2010;Suelter 1967;Suelter et al 1966;Yu et al 2003). Kinetic data Feng Li and Ting Yu equally contribute to this work.…”

Section: Introductionmentioning

confidence: 99%

“…Changes in the tryptophan residues of the solvating environment could indirectly reflect the protein's conformational changes (Kayne and Suelter 1965;Suelter 1967). Three tryptophan residues are present per PK monomer: Trp-157 is in domain B and close to the active site, whereas Trp-481 and Trp-514 are in domain C and close to the Y-interface (Larsen et al 1997;Muirhead et al 1986).…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Probing the catalytic allosteric mechanism of rabbit muscle pyruvate kinase by tryptophan fluorescence quenching

Zhao

et al. 2012

Eur Biophys J

View full text Add to dashboard Cite

Pyruvate kinase acts as an allosteric enzyme, playing a crucial role in the catalysis of the final step of the glycolytic pathway. In this study, site-specific mutagenesis and tryptophan fluorescence quenching were used to probe the catalytic allosteric mechanism of rabbit muscle pyruvate kinase. Movement of the B domain was found to be essential for the catalytic reaction. Rotation of the B domain in the opening of the cleft between domains B and A induced by the binding of activating cations allows substrates to bind, whereas substrate binding shifts the rotation of the B domain in the closure of the cleft. Trp-157 accounts for the differences in tryptophan fluorescence signal with and without activating cations and substrates. Trp-481 and Trp-514 are brought into an aqueous environment after phenylalanine binding.

show abstract

Section: Interaction With Activating Cationscontrasting

confidence: 50%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Probing the catalytic allosteric mechanism of rabbit muscle pyruvate kinase by tryptophan fluorescence quenching

Zhao

et al. 2012

Eur Biophys J

View full text Add to dashboard Cite

show abstract

“…The emission spectra obtained when the enzyme was excited at 280, 292, and 297 nm were recorded to yield the same fluorescence at 375 nm. This procedure normalizes the spectra to give the same tryptophanyl fluorescence (15). Under these conditions, the difference between the emission spectrum obtained at 280 nm excitation and that obtained at 292 nm excitation represents tyrosyl fluorescence, which has a peak at 303 nm (Fig.…”

Section: Methodsmentioning

confidence: 99%

Photoinactivation of Detergent-Solubilized Plasma Membrane ATPase from Rosa damascena

Imbrie

Murphy²

1984

Plant Physiol.

View full text Add to dashboard Cite

The photochemistry of vesicular and detergent-solubilized preparations of plasma membrane-associated ATPase was investigated in Rosa damascena. The cholate-solubilized ATPase activity fractionated into two peaks on a Sephadex G-150 column with simple, but different ultraviolet (UV) sensitivities. The larger enzyme was UV sensitive; the smaller enzyme was relatively insensitive. The activity of both ATPase fractions depended on environment: both were inactive in cholate, relatively inactive in phosphatidylcholine, phosphatidylethanolamine and phosphatidylinositol, and active in phosphatidylglycerol and phosphatidylserine. The UV sensitivities of both fractions also depended on their environment. For the UV sensitive fraction, the action spectrum differed in the 300 to 400 nanometers range when the fraction was irradiated with and without lipids. For the resistant fraction, UV sensitivity at 290 nanometers differed (up to 6-fold) in different lipids. The resistant fraction solubilized in octylglucoside had an action spectrum very different from that in cholate or in lipid vesicles. The absorption spectra of the different preparations reflected the action spectra. For both UV sensitive and insensitive fractions, the action spectra for photoinactivation had peaks at 290 nanometers, suggesting that the chromophores were tryptophanyl residues. The loss of ATPase activity was strictly correlated with the loss of fluorescence from tryptophan in the partially purified enzymes. Cs' protected the UV sensitive activity but not the insensitive one. We propose a model which explains the difference in UV sensitivities based on the positions of the tryptophan residues in the two proteins.The inactivation of a Mg2"-requiring, K+- MATERIALS AND METHODSPlant materials used and procedures for detergent solubilization and gel filtration, as well as ATPase and protein assays used, were described previously (9). Polar Lipid Dispersion. PC,2 PE, and PI from soybean, PG from egg lecithin, and PS from bovine brain, all 99.9% purity, were obtained from Sigma Chemical Company. A mixture of lipids (approximately 50% PC from soybean) was also obtained from Sigma. The lipids were dispersed as described before (9)

show abstract

“…Indeed, several physical studies showed that the conformation of the protein is modified by monovalent cations. Alkali metal ions change the UV, fluorescence polarization, and circular dichroism spectra of the enzyme (7)(8)(9)(10). They also alter its immunoelectrophoretic behavior (11) and the sedimentation velocity of the protein (12).…”

mentioning

confidence: 99%

Pyruvate Kinase Revisited

Oria-Hernández

Cabrera

Pérez‐Montfort

et al. 2005

Journal of Biological Chemistry

View full text Add to dashboard Cite

Effects of Temperature and Activating Cations on the Fluorescence of Pyruvate Kinase^*

Cited by 41 publications

References 10 publications

Probing the catalytic allosteric mechanism of rabbit muscle pyruvate kinase by tryptophan fluorescence quenching

Probing the catalytic allosteric mechanism of rabbit muscle pyruvate kinase by tryptophan fluorescence quenching

Photoinactivation of Detergent-Solubilized Plasma Membrane ATPase from Rosa damascena

Pyruvate Kinase Revisited

Contact Info

Product

Resources

About

Effects of Temperature and Activating Cations on the Fluorescence of Pyruvate Kinase*

Cited by 41 publications

References 10 publications

Probing the catalytic allosteric mechanism of rabbit muscle pyruvate kinase by tryptophan fluorescence quenching

Probing the catalytic allosteric mechanism of rabbit muscle pyruvate kinase by tryptophan fluorescence quenching

Photoinactivation of Detergent-Solubilized Plasma Membrane ATPase from Rosa damascena

Pyruvate Kinase Revisited

Contact Info

Product

Resources

About

Effects of Temperature and Activating Cations on the Fluorescence of Pyruvate Kinase^*