Human peptidylglycine alpha-amidating monooxygenase: cDNA, cloning and functional expression of a truncated form in COS cells.
We have identified a cDNA encoding human peptidylglycine alpha-amidating monooxygenase (PAM; EC 1.14.17.3) with a total length of 3748 bp by screening of a human thyroid carcinoma lambda gt11 library using two heterologous oligonucleotides to conserved regions which derived from frog skin and bovine pituitary PAM sequences. Furthermore we have identified a sequence which differs in a 321 bp deletion. COS cells transfected with a truncated form of this cDNA (lacking the putative carboxyl-terminal transmembrane domain) generated a functional PAM that showed a 20-fold increase of the activity compared to the control and was visualized by immunoblotting.
Glauder
J
J
Universität Frankfurt, Institut für Organische Chemie, FRG.
Ragg
H
H
Rauch
J
J
Engels
J W
JW
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D004273:DNA, Neoplasm / Q000235:genetics*; D015870:Gene Expression; D015723:Gene Library; D005822:Genetic Vectors; D006801:Humans; D006899:Mixed Function Oxygenases; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes; D015345:Oligonucleotide Probes; D000587:Oxidoreductases Acting on CH-NH Group Donors / Q000302:isolation & purification / Q000378:metabolism*; D015183:Restriction Mapping; D013964:Thyroid Neoplasms; D014162:Transfection; D014407:Tumor Cells, Cultured / Q000201:enzymology
D004273:DNA, Neoplasm; D009097:Multienzyme Complexes; D015345:Oligonucleotide Probes; D006899:Mixed Function Oxygenases; C035189:peptidylglycine monooxygenase; D000587:Oxidoreductases Acting on CH-NH Group Donors