LIMITED PROTEOLYSIS OF SNAKE MUSCLE FRUCTOSE 1, 6-BISPHOSPHATASE

Limited proteolysis of snake muscle fructose 1,6-bisphosphatase (EC. 3. 1. 3. 11) with subtilisin resulted in a larger fragment of 29,000 daltons and a smaller fragment of 7000 daltons, and limited tryptic digestion for fructose 1, 6-bisphosphatase mainly produced a 32,000 daltons fragment. Both of the modified enzymes have a 4 to 5 fold increase in activity at pH 9.2 as well as a twofold increase in activity at pH 7.5. At 6μM of AMP more than 90% of the activity was inhibited for the native enzyme but not for the modified enzymes. In case of subtilisin, only 50% of activity was inhibited and in case of trypsin, non inhibition was observed. Results indicated the cleavage sites for the two proteases were apparently different.The above results and the fact, as reported previeusly, that snake muscle fructose 1, 6-bisphosphatase modified with 5, 5'-dithiobis(2-nitrobenzoic acid) also caused a twofold increase in its activity lead us to propose that the catalytic site of snake muscle fructose 1, 6-bis-phosp