Effect of saposins on acid sphingomyelinase.
The effect of saposins (A, B, C and D) on acid sphingomyelinase activity was determined using a crude human kidney sphingomyelinase preparation and a purified sphingomyelinase preparation from human placenta. Saposin D stimulated the activity of the crude enzyme by increasing its apparent Km and Vmax. values for sphingomyelin hydrolysis. Unlike the crude enzyme, the activity of the purified enzyme was strongly inhibited by saposin D as well as other saposins. Saposin D decreased the apparent Km and Vmax values of purified sphingomyelinase activity. The effects of saposin D on the activity of different sphingomyelinase preparations appear to depend on Triton X-100, which is present in the crude enzyme but not in the purified enzyme. When the detergent was removed from the crude preparation, the effect of saposin D changed from being stimulatory to inhibitory. Conversely, when the detergent is added to the purified enzyme, the effect of saposin D on sphingomyelinase activity changed from being inhibitory to stimulatory. While other saposins were inhibitory or had no effect on sphingomyelinase activity in the above assay system, not only saposin D but also saposins A and C exhibited a stimulatory effect upon purified sphingomyelinase activity when the substrate, sphingomyelin, was added in the form of liposomes without detergent. Saposin B was not only inhibitory in the liposome system, but also reduced the stimulatory effect of saposins A, C and D. These observations indicate that the stimulatory effect of saposins A, C and D on acid sphingomyelinase activity is greatly influenced by the physical environment of the enzyme and suggest that similar effects by saposins may be exerted in lysosomal membranes.
Tayama
M
M
Department of Neurosciences, University of California, San Diego, School of Medicine, La Jolla 92093-0634.
Soeda
S
S
Kishimoto
Y
Y
Martin
B M
BM
Callahan
J W
JW
Hiraiwa
M
M
O'Brien
J S
JS
D003902:Detergents; D006023:Glycoproteins / Q000494:pharmacology*; D006801:Humans; D007668:Kidney / Q000201:enzymology; D017830:Octoxynol; D011092:Polyethylene Glycols; D049231:Saposins; D013108:Sphingomyelin Phosphodiesterase / Q000378:metabolism*; D013109:Sphingomyelins / Q000378:metabolism
D003902:Detergents; D006023:Glycoproteins; D049231:Saposins; D013109:Sphingomyelins; D011092:Polyethylene Glycols; D017830:Octoxynol; D013108:Sphingomyelin Phosphodiesterase