The effect of pH on cathepsin activities in mouse liver heterolysosomes.
1. Mouse liver heterolysosomes containing (125)I-labelled albumin were incubated at 35 degrees C in 0.25m-sucrose-0.05m-mercaptoethanol, and various concentrations of buffers at pH4, 5, 7 or 8, and the degradation of intraparticulate protein was measured. 2. Buffers at pH4, 7 or 8 inhibited proteolytic activity and the inhibitions were greater with increasing concentrations of buffers. Tris-acetate buffer, pH8, was a more effective inhibitor. Tris-acetate buffer, pH5, neither inhibited nor stimulated proteolytic activity at concentrations up to 0.1m. 3. Inhibition by pH8 buffers could be accounted for by increased breakage rates of heterolysosomes. 4. Preincubation of heterolysosomes in 0.2m-sodium bicarbonate inhibited proteolytic activity when the particles were washed free of bicarbonate, but the inhibition was reversed if these particles were incubated in media containing pH5 buffer. The inhibition was shown not to be due to an increased breakage of heterolysosomes. 5. It was concluded that the pH within heterolysosomes is about 5, and it is possible to alter reversibly the pH within these particles. The possibility for the existence of an intralysosomal buffering system which maintains the pH at about 5 in heterolysosomes is discussed. This buffering system may be related to the presence of large quantities of acidic lipoproteins in lysosomes observed by other investigators.
D000085:Acetates / Q000378:metabolism; D000418:Albumins / Q000378:metabolism; D000818:Animals; D001639:Bicarbonates / Q000378:metabolism; D002021:Buffers; D002403:Cathepsins / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D007456:Iodine Isotopes; D007668:Kidney / Q000201:enzymology; D008074:Lipoproteins / Q000378:metabolism; D008099:Liver / Q000201:enzymology*; D008247:Lysosomes / Q000201:enzymology*; D008297:Male; D051379:Mice
D000085:Acetates; D000418:Albumins; D001639:Bicarbonates; D002021:Buffers; D007456:Iodine Isotopes; D008074:Lipoproteins; D002403:Cathepsins