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PMID: 192194 Published · ppublish English Journal Article

Purification of glycolytic enzymes by using affinity-elution chromatography.

The Biochemical journal ·Vol. 161 ·No. 2 ·1977-02-01 ·Pages 253-63

Scopes RK

Abstract

1. A systematic procedure for the purification of enzymes by affinity-elution chromatography is described. Enzymes are adsorbed on a cation-exchanger, and eluted with ligands specific for the enzyme concerned. 2. All of the glycolytic and some related enzymes present in rabbit muscle can be purified by the affinity-elution technique. The pH range for adsorption and elution of each enzyme was found, and the effects of minor variations of conditions are described. 3. A description of experimental conditions suitable for affinity elution of each enzyme is given, together with special features relevant to each individual enzyme. 4. Theoretical considerations of affinity elution chromatography are discussed, including its limitations, advantages and disadvantages compared with affinity-adsorption chromatography. Possible developments are suggested to cover enzymes which because of their adsorption characteristics are not at present amenable to affinity-elution procedures.

MeSH Terms
Adenylate Kinase/isolation & purification Animals Chromatography, Affinity/methods Creatine Kinase/isolation & purification Enzymes/isolation & purification Fructose-Bisphosphate Aldolase/isolation & purification Glucose-6-Phosphate Isomerase/isolation & purification Glyceraldehyde-3-Phosphate Dehydrogenases/isolation & purification Glycolysis L-Lactate Dehydrogenase/isolation & purification Muscles/enzymology Phosphofructokinase-1/isolation & purification Phosphoglucomutase/isolation & purification Phosphoglycerate Kinase/isolation & purification Phosphoglycerate Mutase/isolation & purification Phosphopyruvate Hydratase/isolation & purification Phosphorylases/isolation & purification Pyruvate Kinase/isolation & purification Rabbits Triose-Phosphate Isomerase/isolation & purification
Chemicals
Enzymes L-Lactate Dehydrogenase Glyceraldehyde-3-Phosphate Dehydrogenases Phosphorylases Phosphofructokinase-1 Pyruvate Kinase Phosphoglycerate Kinase Creatine Kinase Adenylate Kinase Fructose-Bisphosphate Aldolase Phosphopyruvate Hydratase Triose-Phosphate Isomerase Glucose-6-Phosphate Isomerase Phosphoglycerate Mutase Phosphoglucomutase
Authors & Affiliations
1 authors, click to expand affiliations / ORCID
Scopes R K
References (28)
28 references, click to expand
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj1610253
Published
1977-02-01
Pages
253-63
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1164503
Subset
IM
External Links
PubMed source DOI: 10.1042/bj1610253 PMC Full Text View journal details
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