Hydrolysis by plasma kallikrein of fluorogenic peptides derived from prorenin processing site.
Human plasma kallikrein (HPK) activates plasma prorenin to renin, and the physiological significance of this activation is still unknown. In this paper we investigated the efficiency and the cleavage pattern of the hydrolysis by HPK of the internally quenched fluorescent peptides (qf-peptides) derived from the amino acid sequence of human prorenin cleavage site. The peptide Abz-F-S-Q-P-M-K-R-L-T-L-G-N-T-T-Q-EDDnp (Abz=ortho-aminobenzoic acid, and EDDnp=N-[2,4-dinitrophenyl]-ethylene diamine), that corresponds to the amino acid sequence P(7) to P(7)' of human prorenin cleavage site, is hydrolyzed at the correct processing site (R-L bond) with k(cat)/K(m)=85 mM(-1) s(-1). Alanine was scanned in all positions from P(5) to P(5)' in order to investigate the substrate specificity requirements of HPK. The qf-peptides derived from the equivalent segment of rat prorenin, that has Lys-Lys as basic amino acid pair, and the peptide Abz-NVTSPVQ-EDDnp that contains the proposed cleavage site of rat prorenin have very low susceptibility to hydrolysis by rat plasma kallikrein. These data are according to the previously reported absence of rat plasma prorenin activation by rat plasma kallikrein (RPK), and with the view that prorenin activation in rat requires alternative enzymes and/or mechanism. All the obtained peptides described in this paper were also assayed with bovine trypsin that was taken as a reference protease because it is commonly used to activate prorenin.
Almeida
P C
PC
Department of Biophysics, Escola Paulista de Medicina, Rua Tres de Maio, 100, 04044-020, São Paulo, Brazil.
Chagas
J R
JR
Cezari
M H
MH
Juliano
M A
MA
Juliano
L
L
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D004792:Enzyme Precursors / Q000737:chemistry*; D006801:Humans; D006868:Hydrolysis; D007610:Kallikreins / Q000097:blood*; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D051381:Rats; D012083:Renin / Q000737:chemistry*; D017386:Sequence Homology, Amino Acid
D004792:Enzyme Precursors; D010455:Peptides; D007610:Kallikreins; D012083:Renin