Investigations on the release of membrane‐bound glyceral‐dehyde‐3‐phosphate dehydrogenase

Investigations on the binding of soluble GAPDH to the membrane fraction of rabbit reticulocytes

“…The enzyme binding to the membranes is very firm. As already reported [3,4], it is possible to release the membrane-bound GAPDH of rabbit reticulocytes by different procedures. The reversibility of enzyme release and binding is a necessary condition for a possible regulatory importance of an enzyme binding.…”

“…In spite of this free capacity only 42% of the added enzyme activity is rebound. This incomplete rebinding is contradictory to the high enzyme affinity of the membrane fraction shown in repeated washing procedures [3]. The addition of more GAPDH causes an increase of the bound enzyme activity, which is higher than the proportional one (compare the percentages in table 1).…”

“…The starting material (bound GAPDH) was prepared by hypotonic hemolysis of rabbit reticulocytes [3]. Soluble GAPDH was obtained by incubation of bound GAPDH in 100 mM KC1 containing 10 mM Tris (pH 8.2) for 10 min.…”

Fluorescent probe studies on binding of glyceraldehyde‐3‐phosphate dehydrogenase to phosphatidylinositol liposomes Further evidence for conformational changes

“…Its association with membranes is affected by the environmental parameters such as: pH, ionic strength, ionic metabolites and detergents [7][8][9]. It indicates that the binding is controlled by nonspecific, electrostatic interactions.…”

“…It is a well established fact that glyceraldehyde-3-phosphate dehydrogenase belongs to peripheral membrane proteins [1][2][3][4][5][6]. Its association with membranes is affected by the environmental parameters such as: pH, ionic strength, ionic metabolites and detergents [7][8][9]. It indicates that the binding is controlled by nonspecific, electrostatic interactions.…”