Characterization of a tissue kallikrein inhibitor isolated from Bauhinia bauhinioides seeds: inhibition of the hydrolysis of kininogen related substrates.
Trypsin inhibitors were purified from a saline extract of Bauhinia bauhinioides seeds by ion-exchange column chromatography on DEAE-Sephadex, gel filtration on Superose 12 column, Mono Q ion-exchange chromatography or, alternatively, by affinity chromatography on trypsin-Sepharose. Both B. bauhinioides isolated inhibitors, BbTI-I and BbTI-II, inhibit trypsin being the dissociation constant 0.6 and 0.36 nM, respectively. BbTI-II only inhibits porcine pancreatic kallikrein hydrolysis of H-Pro-Phe-Arg-AMC (Ki 2.0 nM); the bradykinin-containing sequence LGMISLMKRPPGFSPFRSSRI-NH2 and the two kininogen related flanking quenched substrates Abz-MISLMKRP-EDDnp (Ki 2.0 nM) and Abz-FRSSRQ-EDDnp (Ki 2.5 nM).
Oliva
M L
ML
Departamento de Bioquímica, UNIFESP, Escola Paulista de Medicina, São Paulo, SP, Brazil.
Mendes
C R
CR
Juliano
M A
MA
Chagas
J R
JR
Rosa
J C
JC
Greene
L J
LJ
Sampaio
M U
MU
Sampaio
C A
CA
D000595:Amino Acid Sequence; D007887:Fabaceae / Q000737:chemistry*; D006868:Hydrolysis / Q000187:drug effects; D007704:Kininogens / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000737:chemistry* / Q000302:isolation & purification; D010946:Plants, Medicinal; D012639:Seeds / Q000737:chemistry; D017386:Sequence Homology, Amino Acid; D015842:Serine Proteinase Inhibitors / Q000737:chemistry* / Q000302:isolation & purification; D013379:Substrate Specificity; D020840:Tissue Kallikreins / Q000037:antagonists & inhibitors*
D007704:Kininogens; D010940:Plant Proteins; D015842:Serine Proteinase Inhibitors; D020840:Tissue Kallikreins