Aggregation of the inactive form of human alpha-galactosidase in the endoplasmic reticulum

35S-Labeled mutant alpha-galactosidases (Q279E and R301Q) expressed in COS1 cells were detected as two forms (46-kDa protein and gel-top aggregate) on SDS-polyacrylamide gel electrophoresis under nonreducing conditions. The 46-kDa protein disappeared rapidly, but the aggregate decreased slowly. The accumulation of aggregate was observed in COS1 cells expressing either Q279E or R301Q on Western blot analysis. The aggregate was mainly recovered in the fraction extracted with 1% Triton X-100 and had no catalytic activity. The COS1 cells expressing Q279E were treated with 10 microgram/ml brefeldin A or 5 microM monensin. Treatment with brefeldin A caused a decrease in the alpha-galactosidase activity and an increase in the amount of aggregate, but the amount of aggregate did not change on monensin treatment.