Role of antiproteolytic heparin-binding serum protein(s) in modulating the levels of sialyl- and galactosyltransferase activity released during the incubation of rat jejunal slices.
1. Sialyltransferase released into the medium during the incubation of rat jejunal slices in serum-free buffer, was susceptible to proteolytic degradation. Heat inactivated horse serum or its antiproteolytic heparin-binding fraction was found to be necessary in determining the activity of sialyltransferase released (Nadkarni et al., 1991). 2. In the present study, we have shown that heat inactivated rat serum (HRS) or its antiproteolytic heparin-binding fraction (HBF) had a role in determining the sialyltransferase activity released during jejunal slice incubations. 3. Galactosyltransferase was also released during incubations, but was not proteolytically degraded and the presence of HRS or HBF in incubations did not alter the levels of galactosyltransferase activity released. 4. Trypsin activity in serum-free incubation medium was higher compared to medium containing HRS. 5. Addition of serum-free medium obtained from 4 hr incubations of the jejunal slices, to medium obtained from parallel incubations done in the presence of HRS, caused inhibition of sialyl- but not galactosyltransferase activity. 6. In jejunal homogenates stored at -20 degrees C, sialyltransferase activity was decreased during 0-45 days of storage, whereas galactosyltransferase activity remained fairly stable for upto 56 days. 7. Inclusion of HRS or HBF in homogenates resulted in higher sialyl- but not galactosyltransferase activity compared to serum-free homogenate samples. 8. The results suggest that HRS or its antiproteolytic heparin-binding proteins have a role in determining the sialyltransferase activity released from the jejunal slices. In contrast galactosyltransferase released was not susceptible to proteolysis, and HRS or HBF was not required to express its activity.
Nadkarni
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Department of Biochemistry, Memorial University, St. John's, Newfoundland, Canada.
Mookerjea
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S
D000818:Animals; D001798:Blood Proteins / Q000494:pharmacology*; D002352:Carrier Proteins / Q000494:pharmacology*; D005700:Galactosyltransferases / Q000378:metabolism*; D006493:Heparin / Q000378:metabolism* / Q000494:pharmacology; D066298:In Vitro Techniques; D007583:Jejunum / Q000201:enzymology*; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D012799:Sialyltransferases / Q000378:metabolism*; D014357:Trypsin / Q000378:metabolism
D001798:Blood Proteins; D002352:Carrier Proteins; D006493:Heparin; D005700:Galactosyltransferases; D012799:Sialyltransferases; D014357:Trypsin