Puromycin insensitive leucyl-specific aminopeptidase (PILSAP) is involved in the activation of endothelial integrins.
We previously reported that mouse orthologue of puromycin insensitive leucyl-specific aminopeptidase (mPILSAP) played an important role in angiogenesis by regulating the proliferation and migration of endothelial cells (ECs) (Miyashita et al., 2002. Blood 99:3241-3249). Here, we examined the mechanism as to how mPILSAP regulates the migration of ECs. Cell adhesion through integrins plays a crucial role in cell migration, and ECs use at least type-1 collagen receptor integrin alpha2beta1, fibronectin receptor alpha5beta1, and vitronectin receptors alphavbeta3 and alphavbeta5. mPILSAP antisense oligodeoxynucleotide (AS-ODN) or leucinethiol (LT), a leucyl-aminopeptidase inhibitor, did not affect the attachment but did significantly inhibit the spreading of cells of the murine endothelial cell line MSS31 when they were plated on vitronectin-, fibronectin-, or type-1 collagen, although they did not affect the expression of integrin alpha2, alpha5, alphav, beta1, beta3, and beta5 subunits on the cell surface. AS-ODN and LT also inhibited the tyrosine phosphorylation of FAK when cells were plated on vitronectin, fibronectin, or type-1 collagen. This inhibition of cell spreading and of tyrosine phosphorylation of FAK could be negated by Mg(2+). These results suggest that mPILSAP is involved in the activation of endothelial integrins.
Akada
Tetsuya
T
Department of Vascular Biology, Institute of Development, Aging and Cancer, Tohoku University, Sendai, Japan.
Yamazaki
Tohru
T
Miyashita
Hiroki
H
Niizeki
Osamu
O
Abe
Mayumi
M
Sato
Akira
A
Satomi
Susumu
S
Sato
Yasufumi
Y
D000730:Androstadienes / Q000494:pharmacology; D000818:Animals; D015815:Cell Adhesion Molecules / Q000378:metabolism; D002461:Cell Line, Transformed; D002465:Cell Movement / Q000187:drug effects; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology / Q000201:enzymology* / Q000502:physiology; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology; D005109:Extracellular Matrix / Q000737:chemistry / Q000378:metabolism; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; D016023:Integrins / Q000737:chemistry / Q000502:physiology*; D007930:Leucine / Q000031:analogs & derivatives / Q000737:chemistry / Q000502:physiology; D007931:Leucyl Aminopeptidase / Q000037:antagonists & inhibitors* / Q000378:metabolism / Q000502:physiology*; D008274:Magnesium / Q000494:pharmacology; D051379:Mice; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D000081082:Phosphoinositide-3 Kinase Inhibitors; D010766:Phosphorylation / Q000187:drug effects; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology*; D011505:Protein-Tyrosine Kinases / Q000187:drug effects / Q000378:metabolism; D011691:Puromycin / Q000494:pharmacology*; D038981:Receptors, Collagen / Q000378:metabolism; D017469:Receptors, Fibronectin / Q000378:metabolism; D019030:Receptors, Vitronectin / Q000378:metabolism; D013438:Sulfhydryl Compounds; D000077191:Wortmannin
D000730:Androstadienes; D015815:Cell Adhesion Molecules; D004791:Enzyme Inhibitors; D016023:Integrins; D016376:Oligonucleotides, Antisense; D000081082:Phosphoinositide-3 Kinase Inhibitors; D011500:Protein Synthesis Inhibitors; D038981:Receptors, Collagen; D017469:Receptors, Fibronectin; D019030:Receptors, Vitronectin; D013438:Sulfhydryl Compounds; C468280:leucinethiol; D011691:Puromycin; D011505:Protein-Tyrosine Kinases; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; C496051:Ptk2 protein, mouse; D007931:Leucyl Aminopeptidase; D007930:Leucine; D008274:Magnesium; D000077191:Wortmannin