Some kinetic and molecular properties of yeast phosphofructokinase

Kopperschläger, Gèrhard; Freyer, R; Diezel, W; Hofmann, E

doi:10.1016/0014-5793(68)80041-9

Cited by 66 publications

(20 citation statements)

References 12 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Since the speci®c interaction between phosphofructokinase and Cibacron blue F3GA (CB) was ®rst reported (Kopperschlager et al, 1968), dyeÀligand chromatography has been recognized as a powerful technique for protein puri®cation (Arica et al, 1998;Clonis, 1988;Lowe et al, 1981;McCreath et al, 1995). For largescale anity chromatography, the triazine dye CB linked to agarose beads or other matrices has remarkable advantages over conventional anityÀligand systems .…”

Section: Introductionmentioning

confidence: 99%

Further studies on the contribution of electrostatic and hydrophobic interactions to protein adsorption on dye–ligand adsorbents

Zhang

Sun

2001

Biotech & Bioengineering

View full text Add to dashboard Cite

The adsorption equilibria of bovine serum albumin (BSA), gamma-globulin, and lysozyme to three kinds of Cibacron blue 3GA (CB)-modified agarose gels, 6% agarose gel-coated steel heads (6AS), Sepharose CL-6B, and a home-made 4% agarose gel (4AB), were studied. We show that ionic strength has irregular effects on BSA adsorption to the CB-modified affinity gels by affecting the interactions between the negatively charged protein and CB as well as CB and the support matrix. At low salt concentrations, the increase in ionic strength decreases the electrostatic repulsion between negatively charged BSA and the negatively charged gel surfaces, thus resulting in the increase of BSA adsorption. This tendency depends on the pore size of the solid matrix, CB coupling density, and the net negative charges of proteins (or aqueous - phase pH value). Sepharose gel has larger average pore size, so the electrostatic repulsion-effected protein exclusion from the small gel pores is observed only for the affinity adsorbent with high CB coupling density (15.4 micromol/mL) at very low ionic strength (NaCl concentration below 0.05 M in 10 mM Tris-HCl buffer, pH 7.5). However, because CB-6AS and CB-4AB have a smaller pore size, the electrostatic exclusion effect can be found at NaCl concentrations of up to 0.2 M. The electrostatic exclusion effect is even found for CB-6AS with a CB density as low as 2.38 micromol/mL. Moreover, the electrostatic exclusion effect decreases with decreasing aqueous-phase pH due to the decrease of the net negative charges of the protein. For gamma-globulin and lysozyme with higher isoelectric points than BSA, the electrostatic exclusion effect is not observed. At higher ionic strength, protein adsorption to the CB-modified adsorbents decreases with increasing ionic strength. It is concluded that the hydrophobic interaction between CB molecules and the support matrix increases with increasing ionic strength, leading to the decrease of ligand density accessible to proteins, and then the decrease of protein adsorption. Thus, due to the hybrid effect of electrostatic and hydrophobic interactions, in most cases studied there exists a salt concentration to maximize BSA adsorption.

show abstract

Section: Introductionmentioning

confidence: 99%

Further studies on the contribution of electrostatic and hydrophobic interactions to protein adsorption on dye–ligand adsorbents

Zhang

Sun

2001

Biotech & Bioengineering

View full text Add to dashboard Cite

show abstract

“…Strongly coordinating and specific, interactions like hydrogen bonding, van der Waals force, electrostatic force and hydrophobic interaction [40][41][42][43] are responsible for the molecular recognition [5,13,44] and occur at the natural ligand-binding site of the protein. The dyes offered different characteristics depending upon the cumulative effect of the number of benzene rings (predominant), sulphonate groups, and other ionic groups attached to them despite the fact that all of them have a basic triazine nucleus.…”

Section: Discussionmentioning

confidence: 99%

“…Most of the dye and protein interaction studies have been performed on the blue sepharose (cibacron blue linked to beaded agarose) involving different dehydrogenase [12,13].…”

Section: Introductionmentioning

confidence: 99%

Assessment of Pseudoaffinity Chromatography Using Textile Dyes for Isolation of Buffalo Pituitary Luteinizing Hormone

Arora

Patel

Muralidhar

2012

ISRN Chromatography

View full text Add to dashboard Cite

Extensive investigation has been carried out to elucidate the mechanisms involved in pseudoligand affinity chromatography using textile dyes, and, empirically, it has been attributed to the chemical and steric structures of dye and protein. Possibly, a variety of interactions especially ionic and/or hydrophobic influence with a varying share in the binding and differ from protein to protein and from dye to dye. In this study, we have attempted to understand the effect of various biophysical parameters like the nature of the eluant, pH, and ionic strength on the binding of crude luteinizing hormone (LH) with various triazine-based dyes and thus predict their nature. Based on the elution patterns, cibacron and reactive brown suggested a dual electrostatic and hydrophobic nature. Reactive blue and reactive yellow reflected a major electrostatic/ionic nature with yellow offering 50-fold purification in a single step, while reactive red and reactive green had a predominant hydrophobic nature. Appreciably, reactive red was binding LH very tightly unlike other dyes, and addition of the arginine in the elution buffer substantially weakened the protein-dye interactions. pH was observed to be a principal factor assisting the protein-dye binding as well as hydrophobicity of the dye and the proteins.

show abstract

“…Die Substrat-Geschwindigkeitskurven von I T P und UTP zeigen, wie der Anfangsteil der ATP-Kurve, einen hyperbolischen Kurvenverlauf (Fig.4). Eine Enzymhemmung ist selbst bei hoheren Konzentrationen dieser Phosphatdenatoren nicht nachweisbar [15,20]. Die Michaelis-Konstanten fur die drei genannten Phosphatdonatoren sind in Tabelle 1 wiedergegeben.…”

Section: Abhangigkeit Der Enzymaktivitat Vom Phosphatdonatorunclassified

Hefe‐Phosphofruktokinase

Freyer¹,

Kubel²,

Hofmann³

1970

European Journal of Biochemistry

View full text Add to dashboard Cite

The kinetic behaviour of different preparations of yeast phosphofructokinase is described, predominantly regarding the interactions between substrate-and effector-binding sites.The substrate-binding site for ATP reacts with I T P and UTP too, whereas the ATP-inhibitor site possesses higher specsty: other nucleoside triphosphates do not compete with ATP for this site.Binding of protons to the enzyme produces a decrease in cooperativity towards fructose-6-phosphate with ATP as well as with ITP and UTP as phosphate donors, accompanied by a lowering of the fructose-6-phosphate concentration necessary for half-maximal activity. At the same time the inhibitory action of ATP is descreased.AMP is bound t o an effector site evidently different from the ATP-inhibitor site.With ITP as phosphate donor, AMP shows no activating effect. AMP decreases the apparent K , value for fructose-6-phosphate too without affecting cooperative interactions. Magnesium and ammonium ions are necessary for full enzymatic activity. The latter increases maximal activity and decreases the half-maximal concentration of fructose-6-phosphate. Free ATP seems to be a stronger inhibitor for yeast phosphofructokinase than the magnesium-ATP-chelate.

show abstract

Some kinetic and molecular properties of yeast phosphofructokinase

Cited by 66 publications

References 12 publications

Further studies on the contribution of electrostatic and hydrophobic interactions to protein adsorption on dye–ligand adsorbents

Further studies on the contribution of electrostatic and hydrophobic interactions to protein adsorption on dye–ligand adsorbents

Assessment of Pseudoaffinity Chromatography Using Textile Dyes for Isolation of Buffalo Pituitary Luteinizing Hormone

Hefe‐Phosphofruktokinase

Contact Info

Product

Resources

About