Interaction of rabbit muscle enolase and 3-phosphoglycerate mutase studied by ELISA and by batch gel filtration.
The interaction of rabbit skeletal muscle enolase and 3-phosphoglycerate mutase was detected by an ELISA test, a batch gel-filtration technique, and fluorescence anisotropy measurements, and the activity of enolase was determined to be a function of mutase concentration. The apparent dissociation constant of this enzyme complex is approximately 1 microM. This value seems to be independent of the presence (in fluorescence anisotropy measurements) or the absence (in activity as well as in ELISA experiments) of fluorescein isothiocyanate used widely as a label for determining the complex formation between enzymes in fluorescence anisotropy measurements.
Nazaryan
K B
KB
Institute of Experimental Biology, Armenian Academy of Sciences, Yerevan.
Climent
F
F
Simonian
S
S
Tompa
P
P
Batke
J
J
D000818:Animals; D001921:Brain / Q000201:enzymology; D002417:Cattle; D002850:Chromatography, Gel; D004797:Enzyme-Linked Immunosorbent Assay; D016650:Fluorescein-5-isothiocyanate; D005454:Fluorescence Polarization; D005456:Fluorescent Dyes; D046911:Macromolecular Substances; D009132:Muscles / Q000201:enzymology*; D010736:Phosphoglycerate Mutase / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D010751:Phosphopyruvate Hydratase / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D011817:Rabbits
D005456:Fluorescent Dyes; D046911:Macromolecular Substances; D010751:Phosphopyruvate Hydratase; D010736:Phosphoglycerate Mutase; D016650:Fluorescein-5-isothiocyanate