Direct proton magnetic resonance determination of the pKa of the active center histidine in thiolsubtilisin

The serine proteases constitute a group of endopeptidases whose members owe their catalytic activity to the presence of a catalytic triad of amino acids consisting of a serine, a histidine and an aspartate. The pKa values for this histidine have been determined for several cases in which there is a...

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Veröffentlicht in:	Protein science 2002-04, Vol.11 (4), p.965-973
Hauptverfasser:	Kahyaoglu, Ara, Jordan, Frank
Format:	Artikel
Sprache:	eng
Schlagworte:	Binding Sites catalytic triad Cysteine - metabolism DTNB, 5,5′‐dithiobis(2‐nitrobenzoic acid) DTT, dithiothreitol EDTA, ethylenediaminetetraacetic acid Histidine - chemistry Hydrogen Bonding Hydrogen-Ion Concentration KPi, inorganic phosphate, potassium salt Magnetic Resonance Spectroscopy MIP, monoisopropylphosphoryl‐Ser 195 (trypsin and chymotrypsin) or monoisopropylphosphoryl‐Ser 221 (subtilisins) derivative of the respective serine protease Models, Molecular NMR, nuclear magnetic resonance Protein Conformation protein dielectric constant p‐NPA, p‐nitrophenylacetate Serine Endopeptidases - analysis serine protease Subtilisins - chemistry s‐AAPF‐pNA, succinyl‐(L)Ala‐(L)Ala‐(L) Pro‐(L)Phe‐p‐nitroanilide Thiolsubtilisin
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