A cellulose-binding domain-fused recombinant human T cell connective tissue-activating peptide-III manifests heparanase activity.
The chemokine connective tissue-activating peptide (CTAP)-III, which belongs to the leukocyte-derived growth factor family of mediators, was previously shown to be mitogenic for fibroblasts. However, it has recently been shown that CTAP-III, released from platelets, can act like a heparanase enzyme and degrade heparan sulfate. This suggests that CTAP-III may also function as a proinflammatory mediator. We have successfully cloned CTAP-III from a lambdagt11 cDNA library of PHA-activated human CD4(+) T cells and produced recombinant CTAP-III as a fusion protein with a cellulose-binding domain moiety. This recombinant CTAP-III exhibited heparanase activity and released degradation products from metabolically labeled, naturally produced extracellular matrix. We have also developed polyclonal and monoclonal antibodies, and these antibodies against the recombinant CTAP-III detected the CTAP-III molecule in human T cells, polymorphonuclear leukocytes, and placental extracts. Thus, our study provides tools to examine further immune cell behavior in inflamed sites rich with extracellular moieties and proinflammatory mediators.
Department of Immunology, The Weizmann Institute of Science, Rehovot, 76100, Israel.
M
M
Rechter
O
O
Lider
L
L
Cahalon
E
E
Baharav
M
M
Dekel
D
D
Seigel
I
I
Vlodavsky
H
H
Aingorn
I R
IR
Cohen
O
O
Shoseyov
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001426:Bacterial Proteins / Q000235:genetics*; D015496:CD4-Positive T-Lymphocytes / Q000201:enzymology / Q000378:metabolism*; D002352:Carrier Proteins / Q000235:genetics*; D002482:Cellulose / Q000378:metabolism*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005109:Extracellular Matrix / Q000378:metabolism; D005966:Glucuronidase; D006026:Glycoside Hydrolases / Q000378:metabolism*; D006497:Heparan Sulfate / Q000378:metabolism; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D009504:Neutrophils / Q000201:enzymology / Q000378:metabolism; D010455:Peptides / Q000235:genetics* / Q000378:metabolism; D011233:Precipitin Tests; D011485:Protein Binding / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D000099175:Heparanase
D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D002352:Carrier Proteins; D002482:Cellulose; D005966:Glucuronidase; D006026:Glycoside Hydrolases; D006497:Heparan Sulfate; D010455:Peptides; D011993:Recombinant Fusion Proteins; D000099175:Heparanase; C074074:cellulose binding protein A, Clostridium cellulovorans; C009862:connective tissue-activating peptide