Effect of GM2 activator protein on the enzymatic hydrolysis of phospholipids and sphingomyelin.
GM2 activator protein (GM2AP) is a specific protein cofactor that stimulates the enzymatic hydrolysis of the GalNAc from GM2, a sialic acid containing glycosphingolipid, both in vitro and in lysosomes. While phospholipids together with glycosphingolipids are important membrane constituents, little is known about the possible effect of GM2AP on the hydrolysis of phospholipids. Several recent reports suggest that GM2AP might have functions other than stimulating the conversion of GM2 into GM3 by beta-hexosaminidase A, such as inhibiting the activity of platelet activating factor and enhancing the degradation of phosphatidylcholine by phospholipase D (PLD). We therefore examined the effect of GM2AP on the in vitro hydrolyses of a number of phospholipids and sphingomyelin by microbial (Streptomyces chromofuscus) and plant (cabbage) PLD. GM2AP, at the concentration as low as 1.08 microM (1 microg/50 microl) was found to inhibit about 70% of the hydrolyses of phosphatidylcholine, phosphatidylethanolamine, and phosphatidylinositol by PLD, whereas the same concentration of GM2AP only inhibited about 20-25% of the hydrolysis of sphingomyelin by sphingomyelinase and had no effect on the hydrolysis of sphingosylphosphorylcholine by PLD. Thus, GM2AP exerts strong and broad inhibitory effects on the hydrolysis of phospholipids carried out by plant and microbial PLDs. High ammonium sulfate concentration (1.6 M or 21.1%) masks this inhibitory effect, possibly due to the alteration of the ionic property of GM2AP.
Shimada
Yoshimi
Y
Department of Biochemistry, Tulane University Health Sciences Center School of Medicine, New Orleans, LA 70112, USA.
Li
Yu-Teh
YT
Li
Su-Chen
SC
D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism; D001937:Brassica / Q000201:enzymology; D049289:G(M2) Activator Protein; D005678:G(M2) Ganglioside / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism; D006801:Humans; D006868:Hydrolysis; D051379:Mice; D010739:Phospholipase D / Q000378:metabolism*; D010743:Phospholipids / Q000378:metabolism*; D010940:Plant Proteins / Q000378:metabolism; D011506:Proteins / Q000494:pharmacology / Q000502:physiology*; D049231:Saposins; D049128:Sphingolipid Activator Proteins; D013108:Sphingomyelin Phosphodiesterase / Q000378:metabolism; D013109:Sphingomyelins / Q000378:metabolism*; D013302:Streptomyces / Q000201:enzymology; D010738:Type C Phospholipases / Q000378:metabolism
D001426:Bacterial Proteins; D049289:G(M2) Activator Protein; D006023:Glycoproteins; C483345:PSAP protein, human; D010743:Phospholipids; D010940:Plant Proteins; D011506:Proteins; C483344:Psap protein, mouse; D049231:Saposins; D049128:Sphingolipid Activator Proteins; D013109:Sphingomyelins; D005678:G(M2) Ganglioside; D010738:Type C Phospholipases; D013108:Sphingomyelin Phosphodiesterase; D010739:Phospholipase D