Bovine prostacyclin synthase: purification and isolation of partial cDNA.
Prostacyclin synthase (PGIS) from bovine aorta was purified using conventional purification procedures including detergent solubilization, Sephacryl S-300 gel filtration and Mono Q high performance liquid chromatography. Polyclonal antiserum raised against the 52-kDa protein bound specifically to PGIS. After immobilization to Protein A-sepharose, the PGIS-antibody complex displayed PGIS activity. Results based on (1) elution profile of the Mono Q column, (2) two-dimensional gel electrophoresis and (3) N-terminal amino acid sequence suggested that PGIS is heterogenous. Amino acid sequences of N-terminus and a tryptic peptide led us to isolate a partial cDNA fragment.
Pereira
B
B
Department of Internal Medicine, University of Texas-Houston 77030.
Wu
K K
KK
Wang
L H
LH
D000595:Amino Acid Sequence; D000818:Animals; D001011:Aorta / Q000201:enzymology*; D001483:Base Sequence; D002417:Cattle; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002853:Chromatography, Liquid; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis* / Q000302:isolation & purification* / Q000378:metabolism; D017931:DNA Primers; D018076:DNA, Complementary / Q000302:isolation & purification*; D003902:Detergents; D015180:Electrophoresis, Gel, Two-Dimensional; D019746:Intramolecular Oxidoreductases; D007535:Isomerases / Q000096:biosynthesis* / Q000302:isolation & purification* / Q000378:metabolism; D007700:Kinetics; D008297:Male; D008861:Microsomes / Q000201:enzymology / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009131:Muscle, Smooth, Vascular / Q000201:enzymology*; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D016133:Polymerase Chain Reaction; D044262:Prostaglandin H2; D011463:Prostaglandins H / Q000302:isolation & purification / Q000378:metabolism; D012669:Seminal Vesicles / Q000378:metabolism; D012756:Sheep; D014357:Trypsin
D017931:DNA Primers; D018076:DNA, Complementary; D003902:Detergents; D010446:Peptide Fragments; D011463:Prostaglandins H; D044262:Prostaglandin H2; D003577:Cytochrome P-450 Enzyme System; D014357:Trypsin; D007535:Isomerases; D019746:Intramolecular Oxidoreductases; C022461:prostacyclin synthetase