Structural and inhibitory properties of a plant proteinase inhibitor containing the RGD motif.
Purified from Bauhinia rufa seeds, BrTI is a Kunitz proteinase inhibitor that contains the RGD sequence. BrTI inhibits trypsin (K(iapp) 2.9 nM) and human plasma kallikrein (K(iapp) 14.0 nM) but not other related enzymes. The synthetic peptide YLEPVARGDGGLA-NH(2) (70 microM) inhibited the adhesion to fibronectin of B16F10 (high-metastatic B16 murine mouse melanoma cell line) and of Tm5 (murine melanoma cell lines derived from a non-tumorigenic lineage of pigmented murine melanocytes, melan-a). YLEPVARGEGGLA-NH(2) in which Asp(9) was changed into Glu does not affect the cell attachment. Moreover, this peptide was functional only when the sequence present in the native protein was preserved, since YLIPVARGDGGLA-NH(2) in which Glu(3) was changed into Ile does not interfere with B16F10 and was less effective on Tm5 cell line adhesion. Neither YLEPVARGDGGLA-NH(2), YLIPVARGDGGLA-NH(2) or YLEPVARGEGGLA-NH(2) inhibit the interaction of RAEC (endothelial cell line from rabbit aorta) with fibronectin.
Nakahata
Adriana M
AM
Department of Biochemistry, Universidade Federal de São Paulo-Escola Paulista de Medicina, Rua Três de Maio 100, 04044-020 São Paulo, SP, Brazil.
Bueno
Norlene R
NR
Rocha
Hugo A O
HA
Franco
Célia R C
CR
Chammas
Roger
R
Nakaie
Clovis R
CR
Jasiulionis
Miriam G
MG
Nader
Helena B
HB
Santana
Lucimeire A
LA
Sampaio
Misako U
MU
Oliva
Maria Luiza V
ML
D000595:Amino Acid Sequence; D000818:Animals; D036481:Bauhinia / Q000737:chemistry* / Q000235:genetics; D002448:Cell Adhesion / Q000187:drug effects; D045744:Cell Line, Tumor; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects; D006801:Humans; D066298:In Vitro Techniques; D007610:Kallikreins / Q000037:antagonists & inhibitors; D008546:Melanoma, Experimental / Q000188:drug therapy; D051379:Mice; D008969:Molecular Sequence Data; D009842:Oligopeptides; D010455:Peptides / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology*; D010940:Plant Proteins / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology*; D011817:Rabbits; D017386:Sequence Homology, Amino Acid
C053865:Kunitz-type protease inhibitor, plant; D009842:Oligopeptides; D010455:Peptides; D010940:Plant Proteins; C047981:arginyl-glycyl-aspartic acid; D007610:Kallikreins