Sequence, purification, and cloning of an intracellular serine protease, quiescent cell proline dipeptidase.
We recently observed that specific inhibitors of post-proline cleaving aminodipeptidases cause apoptosis in quiescent lymphocytes in a process independent of CD26/dipeptidyl peptidase IV. These results led to the isolation and cloning of a new protease that we have termed quiescent cell proline dipeptidase (QPP). QPP activity was purified from CD26(-) Jurkat T cells. The protein was identified by labeling with [(3)H]diisopropylfluorophosphate and subjected to tryptic digestion and partial amino acid sequencing. The peptide sequences were used to identify expressed sequence tag clones. The cDNA of QPP contains an open reading frame of 1476 base pairs, coding for a protein of 492 amino acids. The amino acid sequence of QPP reveals similarity with prolylcarboxypeptidase. The putative active site residues serine, aspartic acid, and histidine of QPP show an ordering of the catalytic triad similar to that seen in the post-proline cleaving exopeptidases prolylcarboxypeptidase and CD26/dipeptidyl peptidase IV. The post-proline cleaving activity of QPP has an unusually broad pH range in that it is able to cleave substrate molecules at acidic pH as well as at neutral pH. QPP has also been detected in nonlymphocytic cell lines, indicating that this enzyme activity may play an important role in other tissues as well.
Underwood
R
R
Department of Pathology, Tufts University School of Medicine, Boston, Massachusetts 02111, USA.
Chiravuri
M
M
Lee
H
H
Schmitz
T
T
Kabcenell
A K
AK
Yardley
K
K
Huber
B T
BT
D000595:Amino Acid Sequence; D017209:Apoptosis; D002460:Cell Line; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D004150:Dipeptidases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D018819:Dipeptidyl Peptidase 4 / Q000378:metabolism; D006801:Humans; D019169:Jurkat Cells; D007700:Kinetics; D007963:Leukocytes, Mononuclear / Q000201:enzymology; D008214:Lymphocytes / Q000201:enzymology; D008969:Molecular Sequence Data; D011392:Proline / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D012697:Serine Endopeptidases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D013379:Substrate Specificity
D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins; D011392:Proline; D004150:Dipeptidases; C022449:proline dipeptidase; D018819:Dipeptidyl Peptidase 4; D012697:Serine Endopeptidases