TY - JOUR
T1 - Identification of inhibitors of α2β1 integrin, members of C-lectin type proteins, in Echis sochureki venom
AU - Jakubowski, Piotr
AU - Calvete, Juan J.
AU - Eble, Johannes A.
AU - Lazarovici, Philip
AU - Marcinkiewicz, Cezary
PY - 2013/5/15
Y1 - 2013/5/15
N2 - Snake venom antagonists of α2β1 integrin have been identified as members of a C-lectin type family of proteins (CLP). In the present study, we characterized three new CLPs isolated from Echis sochureki venom, which interact with this integrin. These proteins were purified using a combination of gel filtration, ion exchange chromatography and reverse phase HPLC. Sochicetin-A and sochicetin-B potently inhibited adhesion of cells expressing α2β1[U+F020] integrin and binding of isolated α2β1 [U+F020]ectodomain to collagen I, as well as bound to recombinant GST-α2A domain in ELISA, whereas activity of sochicetin-C in these assays was approximately two orders of magnitude lower. Structurally, sochicetin-B and sochicetin-C are typical heterodimeric αβ CLPs, whereas sochicetin-A exhibits a trimer of its subunits (αβ)3 in the quaternary structure. Immobilized sochicetins supported adhesion of glioma cell lines, LN18 and LBC3, whereas in a soluble form they partially inhibited adhesion of these cells to collagen I. Glioma cells spread very poorly on sochicetin-A, showing no cytoskeleton rearrangement typical for adhesion to collagen I or fibronectin. Adhesion on CLP does not involve focal adhesion elements, such as vinculin. Sochicetin-A also inhibited collagen-induced platelet aggregation, similar to other CLPs' action on the blood coagulation system.
AB - Snake venom antagonists of α2β1 integrin have been identified as members of a C-lectin type family of proteins (CLP). In the present study, we characterized three new CLPs isolated from Echis sochureki venom, which interact with this integrin. These proteins were purified using a combination of gel filtration, ion exchange chromatography and reverse phase HPLC. Sochicetin-A and sochicetin-B potently inhibited adhesion of cells expressing α2β1[U+F020] integrin and binding of isolated α2β1 [U+F020]ectodomain to collagen I, as well as bound to recombinant GST-α2A domain in ELISA, whereas activity of sochicetin-C in these assays was approximately two orders of magnitude lower. Structurally, sochicetin-B and sochicetin-C are typical heterodimeric αβ CLPs, whereas sochicetin-A exhibits a trimer of its subunits (αβ)3 in the quaternary structure. Immobilized sochicetins supported adhesion of glioma cell lines, LN18 and LBC3, whereas in a soluble form they partially inhibited adhesion of these cells to collagen I. Glioma cells spread very poorly on sochicetin-A, showing no cytoskeleton rearrangement typical for adhesion to collagen I or fibronectin. Adhesion on CLP does not involve focal adhesion elements, such as vinculin. Sochicetin-A also inhibited collagen-induced platelet aggregation, similar to other CLPs' action on the blood coagulation system.
KW - C-lectin type proteins
KW - Cell adhesion
KW - Collagen receptors
KW - Glioma cells
KW - Integrins
KW - Snake venom proteins
UR - http://www.scopus.com/inward/record.url?scp=84875662036&partnerID=8YFLogxK
U2 - 10.1016/j.taap.2013.03.002
DO - 10.1016/j.taap.2013.03.002
M3 - ???researchoutput.researchoutputtypes.contributiontojournal.article???
C2 - 23499869
AN - SCOPUS:84875662036
SN - 0041-008X
VL - 269
SP - 34
EP - 42
JO - Toxicology and Applied Pharmacology
JF - Toxicology and Applied Pharmacology
IS - 1
ER -