Novel prolyl tri/tetra-peptidyl aminopeptidase from Streptomyces mobaraensis: substrate specificity and enzyme gene cloning.
The prolyl peptidase that removes the tetra-peptide of pro-transglutaminase was purified from Streptomyces mobaraensis mycelia. The substrate specificity of the enzyme using synthetic peptide substrates showed proline-specific activity with not only tripeptidyl peptidase activity, but also tetrapeptidyl peptidase activity. However, the enzyme had no other exo- and endo-activities. This substrate specificity is different from proline specific peptidases so far reported. The enzyme gene was cloned, based on the direct N-terminal amino acid sequence of the purified enzyme, and the entire nucleotide sequence of the coding region was determined. The deduced amino acid sequence revealed an N-terminal signal peptide sequence (33 amino acids) followed by the mature protein comprising 444 amino acid residues. This enzyme shows no remarkable homology with enzymes belonging to the prolyl oligopeptidase family, but has about 65% identity with three tripeptidyl peptidases from Streptomyces lividans, Streptomyces coelicolor, and Streptomyces avermitilis. Based on its substrate specificity, a new name, "prolyl tri/tetra-peptidyl aminopeptidase," is proposed for the enzyme.
Umezawa
Yukiko
Y
Institute of Life Sciences, Ajinomoto Co., Inc., Kawasaki 210-8681, Japan. yukiko_umezawa@ajinomoto.com
Yokoyama
Keiichi
K
Kikuchi
Yoshimi
Y
Date
Masayo
M
Ito
Kiyoshi
K
Yoshimoto
Tadashi
T
Matsui
Hiroshi
H
D000595:Amino Acid Sequence; D000626:Aminopeptidases; D001483:Base Sequence; D001665:Binding Sites; D002845:Chromatography; D003001:Cloning, Molecular; D004247:DNA / Q000378:metabolism; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000096:biosynthesis* / Q000235:genetics*; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007525:Isoelectric Focusing; D007700:Kinetics; D008969:Molecular Sequence Data; D010455:Peptides; D011392:Proline / Q000737:chemistry; D017386:Sequence Homology, Amino Acid; D057057:Serine Proteases; D013302:Streptomyces / Q000201:enzymology*; D013379:Substrate Specificity; D013696:Temperature; D011503:Transglutaminases / Q000378:metabolism; D000091346:Tripeptidyl-Peptidase 1
D010455:Peptides; D000091346:Tripeptidyl-Peptidase 1; D004247:DNA; D011392:Proline; D011503:Transglutaminases; D010450:Endopeptidases; D057057:Serine Proteases; C501986:prolyl tri-tetra-peptidyl aminopeptidase, Streptomyces mobaraensis; D000626:Aminopeptidases; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases