scholarly journals Inhibition of monkey liver serine hydroxymethyltransferase by Cibacron Blue 3G-A

1980 ◽  
Vol 187 (1) ◽  
pp. 249-252 ◽  
Author(s):  
K S Ramesh ◽  
N Appaji Rao
Keyword(s):  
Absorption Spectrum ◽  
Binding Domain ◽  
Serine Hydroxymethyltransferase ◽  
Free Enzyme ◽  
Cibacron Blue ◽  
Monkey Liver ◽  
Dye Absorption ◽  
Blue Sepharose

Cibacron Blue 3G-A inhibited monkey liver serine hydroxymethyltransferase competitively with respect to tetrahydrofolate and non-competitively with respect to L-serine. NADH, a positive heterotropic effector, failed to protect the enzymes against inhibition by the dye and was unable to desorb the enzyme from Blue Sepharose CL-6B gel matrix. The binding of the dye to the free enzyme was confirmed by changes in the dye absorption spectrum. The results indicate that the dye probably binds at the tetrahydrofolate-binding domain of the enzyme, rather than at the ‘dinucleotide fold’.

scholarly journals Purification and physicochemical, kinetic and immunological properties of allosteric serine hydroxymethyltransferase from monkey liver

1980 ◽  
Vol 187 (3) ◽  
pp. 623-636 ◽  
Author(s):  
K S Ramesh ◽  
N Appaji Rao
Keyword(s):  
Liver Enzyme ◽  
Thermal Inactivation ◽  
Heat Stability ◽  
Serine Hydroxymethyltransferase ◽  
Heat Denaturation ◽  
Monkey Liver ◽  
Catalytically Active ◽  
Antibody Complex ◽  
Bacterial Sources ◽  
Blue Sepharose

The homogeneous serine hydroxymethyltransferase purified from monkey liver, by the use of Blue Sepharose affinity chromatography, exhibited positive homotropic co-operative interactions (h = 2.5) with tetrahydrofolate and heterotropic interactions with L-serine and nicotinamide nucleotides. The enzyme had an unusually high temperature optimum of 60 degrees C and was protected against thermal inactivation by L-serine. The allosteric effects were abolished when the monkey liver enzyme was purified by using a heat-denaturation step in the presence of L-serine, a procedure adopted by earlier workers for the purification of this enzyme from mammalian and bacterial sources. The enzyme activity was inhibited completely by N5-methyltetrahydrofolate, N5-formyltetrahydrofolate, dichloromethotrexate, aminopterin and D-cycloserine, whereas methotrexate and dihydrofolate were partial inhibitors. The insoluble monkey liver enzyme-antibody complex was catalytically active and failed to show positive homotropic co-operative interactions with tetrahydrofolate (h = 1) and heterotropic interactions with NAD+. The enzyme showed a higher heat-stability in a complex with its antibody than as the free enzyme. These results highlight the pitfalls in using a heat-denaturation step in the purification of allosteric enzymes.

scholarly journals The interaction of anthraquinone dyes with the plasmid-mediated OXA-2 β-lactamase

1982 ◽  
Vol 205 (2) ◽  
pp. 413-417 ◽  
Author(s):  
C Monaghan ◽  
S Holland ◽  
J W Dale
Keyword(s):  
Specific Effect ◽  
Nucleotide Binding ◽  
Red Shift ◽  
Side Chain ◽  
Beta Lactamase ◽  
Beta Lactamases ◽  
Cibacron Blue ◽  
Anthraquinone Dyes ◽  
Cibacron Blue 3Ga ◽  
Blue Sepharose

Although beta-lactamases do not require any nucleotide co-substrates, the OXA-2 type is inhibited competitively by Cibacron Blue 3GA, and by other anthraquinone dyes, including some simpler compounds with no side chain. The enzyme causes a red shift in the spectrum of Cibacron Blue. The beta-lactamase can be adsorbed in Blue Sepharose and specifically eluted by benzylpenicillin. These results indicate that the binding of anthraquinone dyes is a specific effect similar to that seen with many nucleotide-binding enzymes.

Purification of human α-2-macroglobulin by chromatography on Cibacron Blue Sepharose

1978 ◽  
Vol 89 (1) ◽  
pp. 274-278 ◽  
Author(s):  
G.D. Virca ◽  
J. Travis ◽  
P.K. Hall ◽  
R.C. Roberts
Keyword(s):  
Cibacron Blue ◽  
Blue Sepharose

Purification of diphtheria toxin by chromatography on Cibacron Blue-Sepharose

1983 ◽  
Vol 39 (8) ◽  
pp. 885-886 ◽  
Author(s):  
G. Antoni ◽  
M. Bigio ◽  
G. Borri ◽  
M. C. Casagli ◽  
P. Neri
Keyword(s):  
Diphtheria Toxin ◽  
Cibacron Blue ◽  
Blue Sepharose

scholarly journals Isolation of albumin from whole human plasma and fractionation of albumin-depleted plasma

1976 ◽  
Vol 157 (2) ◽  
pp. 301-306 ◽  
Author(s):  
J Travis ◽  
J Bowen ◽  
D Tewksbury ◽  
D Johnson ◽  
R Pannell
Keyword(s):  
Ion Exchange ◽  
Human Plasma ◽  
Plasma Proteins ◽  
Deae Cellulose ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Affinity Column ◽  
Plasma Albumin ◽  
Cibacron Blue ◽  
Blue Sepharose

The dye Cibacron Blue F-3-GA was conjugated to Sepharose to provide an affinity column for serum albumin. Passage of whole human plasma through a column of Cibacron Blue-Sepharose results in the removal of approx. 98% of the albumin. The latter can be quantitatively recovered by desorption with NaSCN. Albumin-depleted plasma can be readily resolved into discrete fractions by a combination of conventional biochemical techniques. In particular, the resolution of plasma proteins with properties similar to those of native human plasma albumin can readily be accomplished by ion-exchange chromatography of the Sepharose-dye-treated plasma on DEAE-cellulose.

scholarly journals Regulation of monkey liver serine hydroxymethyltransferase by nicotinamide nucleotide

1978 ◽  
Vol 174 (3) ◽  
pp. 1055-1058 ◽  
Author(s):  
K S Ramesh ◽  
N A Rao
Keyword(s):  
Serine Hydroxymethyltransferase ◽  
Monkey Liver ◽  
First Time ◽  
Metabolic Interconversion ◽  
Time Lead

The positive homotropic binding of tetrahydrofolate to monkey liver serine hydroxymethyltransferase was abolished on preincubating the enzyme with NADH and NADPH. NAD+ was a negative heterotropic effector, whereas NADP+ was without effect. The allosteric effects of nicotinamide nucleotides on the serine hydroxymethyltransferase, reported for the first time, lead to a better understanding of the regulation of the metabolic interconversion of folate coenzymes.

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