Open AccessYeast Inorganic Pyrophosphatase: Studies on Metal Binding
Author(s) -
BAYKOV Alexander A.,
AVAEVA Svetlana M.
Publication year - 1974
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1974.tb03667.x
Subject(s) - chemistry , dissociation constant , pyrophosphate , zinc , inorganic pyrophosphatase , ionic strength , magnesium , binding site , inorganic chemistry , metal , binding constant , hydrolysis , metal ions in aqueous solution , dissociation (chemistry) , pyrophosphatase , enzyme , stereochemistry , biochemistry , aqueous solution , organic chemistry , receptor
A study has been made of the interaction of yeast inorganic pyrophosphatase with two activating cations, Mg 2+ and Zn 2+ , and one inhibitory cation, Ca 2+ at pH 6.5 and ionic strength of unity. These cations were found to partially protect the enzyme against inactivation by iodine. Dissociation constants of enzyme metal complexes estimated from protection studies were 11, 500 and 4000 μM for Zn 2+ , Mg 2+ and Ca 2+ , respectively. An additional, lower affinity site, was found for Zn 2+ ( K D = 3.4 mM). Competition studies showed that magnesium and calcium ions compete for the same site as the tighter bound zinc ion and do not compete for the lower affinity site. Binding of Zn 2+ to both sites is pH‐dependent; the interaction with the high‐affinity site is controlled by a group with p K = 6.6. The binding constant for Mg 2+ is nearly the same as the lower activation constant for this cation in enzymatic hydrolysis of pyrophosphate. Two activating and one inhibitory site were found for Zn 2+ in ATP hydrolysis. The inhibitory site was identified as the lower affinity site deduced from protection studies. Both activation constants for Zn 2+ are higher than the dissociation constant for the tightly bound zinc deduced from protection studies.