Histidine decarboxylase from rat mast cells. Enhanced recovery in cell-free extracts and isotopic labelling.
A procedure for obtaining rat mast-cell histidine decarboxylase in greater than 50% yield in cell-free extracts was developed. The enzyme was found in the supernatant fractions from a 3,500 g and a 105,000 g centrifugation step and was demonstrated to be sensitive to inhibition by alpha-fluoromethylhistidine but not by phenylalanine. Although the enzyme shows a half-life of only 3 h in cell-free extract, the initial high recovery of activity allowed for active-site labelling of the enzyme by [3H]histidine and NaBH4. Labelled protein migrated on non-denaturing polyacrylamide-gradient-gel electrophoresis as a 55,000 Da species.
Feldberg
R S
RS
Department of Biology, Tufts University, Medford, MA 02155.
Iannitti
D A
DA
Cochrane
D E
DE
D000818:Animals; D002262:Carboxy-Lyases / Q000302:isolation & purification*; D002474:Cell-Free System; D004591:Electrophoresis, Polyacrylamide Gel; D006632:Histamine / Q000096:biosynthesis; D006640:Histidine Decarboxylase / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D007424:Intracellular Fluid / Q000201:enzymology; D008297:Male; D008407:Mast Cells / Q000201:enzymology*; D008762:Methylhistidines / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D014316:Tritium
D008762:Methylhistidines; D014316:Tritium; C028309:alpha-fluoromethylhistidine; D006632:Histamine; D002262:Carboxy-Lyases; D006640:Histidine Decarboxylase