Alternagin-C, a nonRGD-disintegrin, induces neutrophil migration via integrin signaling.
Recently, a new protein containing a disintegrin domain, alternagin-C (Alt-C), was purified from Bothrops alternatus venom. Unlike other disintegrins, in Alt-C an ECD amino acid mogif takes the place of the RGD sequence. Most disintegrins contain an RGD/KGD sequence and are very potent inhibitors of platelet aggregation, as well as other cell interactions with the extracellular matrix, including tumor cell metastasis and angiogenesis. The present study investigated the effects of Alt-C on human neutrophil chemotaxis in vitro and the activation of integrin-mediated pathways. Alt-C showed a potent chemotactic effect for human neutrophils when compared to N-formyl-methionyl-leucyl-phenylalanine peptide (fMLP), a classic chemotactic agent. Moreover, preincubation of neutrophils with Alt-C significantly inhibited chemotaxis toward fMLP and itself. In addition, a peptide containing an ECD sequence presented a chemotactic activity and significantly inhibited chemotaxis induced by Alt-C and fMLP. A significant increase of F-actin content was observed in cells treated with Alt-C, showing that the chemotactic activity of Alt-C on neutrophils is driven by actin cytoskeleton dynamic changes. Furthermore, this protein was able to induce an increase of phosphotyrosine content triggering focal adhesion kinase activation and its association with phosphatidylinositol 3-kinase. Alt-C was also able to induce a significant increase in extracellular signal-regulated kinase 2 nuclear translocation. The chemotactic activity of Alt-C was partially inhibited by LY294002, a specific phosphatidylinositol 3-kinase inhibitor, and by PD98056, a Map kinase kinase inhibitor. These findings suggest that Alt-C can trigger human neutrophil chemotaxis modulated by intracellular signals characteristic of integrin-activated pathways and that these effects could be related to the ECD mogif present in disintegrin-like domain.
Mariano-Oliveira
Andréa
A
Departamento de Farmacologia, Instituto de Biologia, Universidade do Estado do Rio de Janeiro, Brazil.
Coelho
Ana Lúcia J
AL
Terruggi
Cristina H B
CH
Selistre-de-Araújo
Heloísa S
HS
Barja-Fidalgo
Christina
C
De Freitas
Marta S
MS
D000199:Actins / Q000378:metabolism; D021581:Active Transport, Cell Nucleus; D020816:Amino Acid Motifs; D000818:Animals; D017837:Bothrops / Q000378:metabolism; D002465:Cell Movement; D002467:Cell Nucleus / Q000378:metabolism; D002633:Chemotaxis; D003599:Cytoskeleton / Q000378:metabolism; D019483:Disintegrins / Q000737:chemistry* / Q000502:physiology*; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005109:Extracellular Matrix / Q000378:metabolism; D005419:Flavonoids / Q000494:pharmacology; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D016023:Integrins / Q000378:metabolism*; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000737:chemistry; D009362:Neoplasm Metastasis; D009389:Neovascularization, Pathologic; D009504:Neutrophils / Q000166:cytology* / Q000378:metabolism; D010455:Peptides / Q000737:chemistry; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D019000:Phosphotyrosine / Q000378:metabolism; D011233:Precipitin Tests; D017434:Protein Structure, Tertiary; D015398:Signal Transduction; D012910:Snake Venoms / Q000378:metabolism; D013997:Time Factors
D000199:Actins; D019483:Disintegrins; D004791:Enzyme Inhibitors; D005419:Flavonoids; D016023:Integrins; D010455:Peptides; D012910:Snake Venoms; C481346:alternagin-C; D019000:Phosphotyrosine; D009240:N-Formylmethionine Leucyl-Phenylalanine; D019869:Phosphatidylinositol 3-Kinases; D019950:Mitogen-Activated Protein Kinase 1; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one