Evidence for an essential histidine in neutral endopeptidase 24.11.
作者： Robert C. Bateman , Louis B. Hersh
DOI: 10.1021/BI00388A009
关键词: Stereochemistry 、 Hydroxylamine 、 Enzyme assay 、 Active site 、 Hydrolysis 、 Enzyme 、 Phosphoramidon 、 Histidine 、 Residue (chemistry) 、 Chemistry
摘要: Rat kidney neutral endopeptidase 24.11, "enkephalinase", was rapidly inactivated by diethyl pyrocarbonate under mildly acidic conditions. The pH dependence of inactivation revealed the modification an essential residue with a pKa 6.1. reaction unprotonated group exhibited second-order rate constant 11.6 M-1 s-1 and accompanied increase in absorbance at 240 nm. Treatment enzyme 50 mM hydroxylamine completely restored activity. These findings indicate histidine pyrocarbonate. Comparison nm single for catalysis. presence this active site indicated (a) protection from provided substrate (b) specific inhibitor phosphoramidon one as determined spectrally. kinetic parameter Vmax/Km upon two residues values 5.9 7.3. It is proposed that having modified participates general acid/base catalysis during hydrolysis 24.11.
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sci-hub.se 参考文章(36)
Edith Wilson Miles, Modification of histidyl residues in proteins by diethylpyrocarbonate. Methods in Enzymology. ,vol. 47, pp. 431- 442 ,(1977) , 10.1016/0076-6879(77)47043-5
M. A. Kerr, A. J. Kenny, The purification and specificity of a neutral endopeptidase from rabbit kidney brush border. Biochemical Journal. ,vol. 137, pp. 477- 488 ,(1974) , 10.1042/BJ1370477
P Dominici, B Tancini, C Borri Voltattorni, Chemical modification of pig kidney 3,4-dihydroxyphenylalanine decarboxylase with diethyl pyrocarbonate. Evidence for an essential histidyl residue. Journal of Biological Chemistry. ,vol. 260, pp. 10583- 10589 ,(1985) , 10.1016/S0021-9258(19)85125-6
D G Jackson, L B Hersh, Reaction of neutral endopeptidase 24.11 (enkephalinase) with arginine reagents. Journal of Biological Chemistry. ,vol. 261, pp. 8649- 8654 ,(1986) , 10.1016/S0021-9258(19)84429-0
J. John Holbrook, V. Ann Ingram, Ionic properties of an essential histidine residue in pig heart lactate dehydrogenase Biochemical Journal. ,vol. 131, pp. 729- 738 ,(1973) , 10.1042/BJ1310729
Christopher M. TOPHAM, Keith DALZIEL, Chemical modification of sheep-liver 6-phosphogluconate dehydrogenase by diethylpyrocarbonate FEBS Journal. ,vol. 155, pp. 87- 94 ,(1986) , 10.1111/J.1432-1033.1986.TB09461.X
L B Hersh, K Morihara, Comparison of the subsite specificity of the mammalian neutral endopeptidase 24.11 (enkephalinase) to the bacterial neutral endopeptidase thermolysin. Journal of Biological Chemistry. ,vol. 261, pp. 6433- 6437 ,(1986) , 10.1016/S0021-9258(19)84580-5
Shigeru KUNUGI, Hideo HIROHARA, Norio ISE, pH and temperature dependences of thermolysin catalysis. Catalytic role of zinc-coordinated water. FEBS Journal. ,vol. 124, pp. 157- 163 ,(1982) , 10.1111/J.1432-1033.1982.TB05919.X
R.B. Harris, I.B. Wilson, Irreversible inhibition of bovine lung angiotensin I-converting enzyme with p-[N,N-bis(chloroethyl)amino]phenylbutyric acid (chlorambucil) and chlorambucyl L-proline and with evidence that an active site carboxyl group is labeled. Journal of Biological Chemistry. ,vol. 257, pp. 811- 815 ,(1982) , 10.1016/S0021-9258(19)68269-4
Anthony J. Turner, Rebecca Matsas, A.John Kenny, Are there neuropeptide-specific peptidases? Biochemical Pharmacology. ,vol. 34, pp. 1347- 1356 ,(1985) , 10.1016/0006-2952(85)90669-0