PMID- 9496431
OWN - NLM
STAT- MEDLINE
DCOM- 19980604
LR  - 20190516
IS  - 0100-879X (Print)
IS  - 0100-879X (Linking)
VI  - 30
IP  - 10
DP  - 1997 Oct
TI  - Specific fluorogenic substrates for neprilysin (neutral endopeptidase, EC 
      3.4.24.11) which are highly resistant to serine- and metalloproteases.
PG  - 1157-62
AB  - Two intramolecularly quenched fluorogenic peptides containing o-aminobenzoyl 
      (Abz) and ethylenediamine 2,4-dinitrophenyl (EDDnp) groups at amino- and 
      carboxyl-terminal amino acid residues, Abz-DArg-Arg-Leu-EDDnp (Abz-DRRL-EDDnp) 
      and Abz-DArg-Arg-Phe-EDDnp (Abz-DRRF-EDDnp), were selectively hydrolyzed by 
      neutral endopeptidase (NEP, enkephalinase, neprilysin, EC 3.4.24.11) at the 
      Arg-Leu and Arg-Phe bonds, respectively. The kinetic parameters for the 
      NEP-catalyzed hydrolysis of Abz-DRRL-EDDnp and Abz-DRRF-EDDnp were K(m) = 2.8 
      microM, kcat = 5.3 min-1, kcat/K(m) = 2 min-1 microM-1 and K(m) = 5.0 microM, 
      kcat = 7.0 min-1, kcat/K(m) = 1.4 min-1 microM-1, respectively. The high 
      specificity of these substrates was demonstrated by their resistance to 
      hydrolysis by metalloproteases [thermolysin (EC 3.4.24.2), angiotensin-converting 
      enzyme (ACE; EC 3.4.24.15)], serineproteases [trypsin (EC 3.4.21.4), 
      alpha-chymotrypsin (EC 3.4.21.1)] and proteases present in tissue homogenates 
      from kidney, lung, brain and testis. The blocked amino- and carboxyl-terminal 
      amino acids protected these substrates against the action of aminopeptidases, 
      carboxypeptidases and ACE. Furthermore, DR amino acids ensured total protection 
      of Abz-DRRL-EDDnp and Abz-DRRF-EDDnp against the action of thermolysin and 
      trypsin. Leu-EDDnp and Phe-EDDnp were resistant to hydrolysis by 
      alpha-chymotrypsin. The high specificity of these substrates suggests their use 
      for specific NEP assays in crude enzyme preparations.
FAU - Medeiros, M A
AU  - Medeiros MA
AD  - Departamento de Fisiologia e Farmacologia, Universidade Federal do Ceara, 
      Fortaleza, Brasil.
FAU - Franca, M S
AU  - Franca MS
FAU - Boileau, G
AU  - Boileau G
FAU - Juliano, L
AU  - Juliano L
FAU - Carvalho, K M
AU  - Carvalho KM
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Brazil
TA  - Braz J Med Biol Res
JT  - Brazilian journal of medical and biological research = Revista brasileira de 
      pesquisas medicas e biologicas
JID - 8112917
RN  - 284SYP0193 (Fluorine)
RN  - EC 3.4.21.- (Serine Endopeptidases)
RN  - EC 3.4.24.- (Metalloendopeptidases)
RN  - EC 3.4.24.11 (Neprilysin)
SB  - IM
MH  - Animals
MH  - Fluorine/metabolism
MH  - Metalloendopeptidases/*metabolism
MH  - Neprilysin/*metabolism
MH  - Rats
MH  - Serine Endopeptidases/*metabolism
EDAT- 1998/03/13 00:00
MHDA- 1998/03/13 00:01
CRDT- 1998/03/13 00:00
PHST- 1998/03/13 00:00 [pubmed]
PHST- 1998/03/13 00:01 [medline]
PHST- 1998/03/13 00:00 [entrez]
AID - 10.1590/s0100-879x1997001000003 [doi]
PST - ppublish
SO  - Braz J Med Biol Res. 1997 Oct;30(10):1157-62. doi: 
      10.1590/s0100-879x1997001000003.