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.
Rechter
M
M
Department of Immunology, The Weizmann Institute of Science, Rehovot, 76100, Israel.
Lider
O
O
Cahalon
L
L
Baharav
E
E
Dekel
M
M
Seigel
D
D
Vlodavsky
I
I
Aingorn
H
H
Cohen
I R
IR
Shoseyov
O
O
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