Purification and cDNA cloning of Xenopus laevis skin peptidylhydroxyglycine N-C lyase, catalyzing the second reaction of C-terminal alpha-amidation.
The alpha-amidation of glycine-extended peptides is a two-step reaction catalyzed by peptidylglycine alpha-hydroxylating monooxygenase (PHM) and peptidylhydroxyglycine N-C lyase (PHL). PHL was purified to homogeneity from Xenopus laevis skin and its partial amino acid sequence (including the N-terminal 35 residues) was determined. It was found that the cDNA codes for a 935-residue precursor protein (AE-III protein), containing the PHM and PHL sequences at its N terminus and C terminus, respectively. The PHM sequence in AE-III protein is completely identical to that deduced from the nucleotide sequence of X. laevis AE-I cDNA, which encodes only PHM, except that the AE-I protein has an extra 10 residues at its C terminus. It is suggested that AE-I and AE-III mRNA are encoded by the same gene and produced by alternative splicing.
Bio-organics Research Department, International Research Laboratories, Ciba-Geigy, Japan.
Y
Y
Iwasaki
T
T
Kawahara
H
H
Shimoi
K
K
Suzuki
O
O
Ghisalba
K
K
Kangawa
H
H
Matsuo
Y
Y
Nishikawa
D000577:Amides / Q000378:metabolism; D019760:Amidine-Lyases; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D002116:Calcitonin / Q000378:metabolism; D002384:Catalysis; D003001:Cloning, Molecular; D004247:DNA / Q000737:chemistry; D004792:Enzyme Precursors / Q000737:chemistry / Q000235:genetics; D008190:Lyases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D012867:Skin / Q000201:enzymology*; D014982:Xenopus laevis
D000577:Amides; D004792:Enzyme Precursors; D002116:Calcitonin; D004247:DNA; D008190:Lyases; D019760:Amidine-Lyases; C068521:peptidylamidoglycolate lyase