Isolierung von Protease-Inhibitoren

Abstract

Unter Berucksichtigung der Gesamtausbeute (79,7 %) an Isomerengemisch ist die von Baer [4Cl bei der Nitroathan-Cyclisierung von 2-0-[(S)-Formyl-methoxy-methyl]-(R)glycerinaldehyd beobachtete CS-Epimerisierung hier auszu-schlieRen. Eingegangen am 30. Juni 1966[ Z 2771

Zur Isolierung von Schweine- und Human-Serumkallikrein durch Affinitätschromatographie. Spezifische Bindung an wasserunlösliche Kunitz-Sojabohnen-Inhibitor-Cellulosen und Dissoziation mit kompetitiven Hemmstoffen (Benzamidin)

“…Seit der Beschreibung der ersten erfolgreichen Isolierungen von Enzyment 10^ und Enzyminhibitoren [11 l durch affmitätschromatographische Methoden haben sich diese Verfahren in einem bemerkenswert kurzen Zeitraum aufgrund ihrer Einfachheit und vielseitigen Anwendbarkeit allgemein durchgesetztt 12 !. Auch die Isolierung der labilen Serumkai likreine wird, wie aus dem in Abb.…”

Affinity Chromatography of Tryptases: Design, Synthesis and Characterization of a Novel Matrix‐Bound Bivalent Inhibitor

“…Stirring was continued overnight before evaporation of the solvent. The obtained oil was dissolved in water (100 mL), acidified by addition of KHSO 4 , and extracted with AcOEt (3 100 mL), and the combined organic phases were washed with brine (1 50 mL) and dried (Na 2 SO 4 ). The product was isolated by precipitation from tert-butyl methyl ether/petroleum ether to give a colorless powder; yield: 1.74 g (70 % over two steps); TLC (cyclohexane/CHCl 3 Z-dl-Phe(3-BocHN-CH 2 )-NHMe (8): Aqueous methylamine (11.85 m, 0.22 mL, 2.63 mmol), HOBt (0.24 g, 1.75 mmol), and EDC (0.50 g, 2.63 mmol) were successively added to an ice-cold (0 8C), stirred solution of 7 (0.75 g, 1.75 mmol) in DMF (30 mL).…”

“…In the late 1960s Fritz et al took advantage of a resin-bound version of the protein trypsin-kallikrein inhibitor for the purification of trypsin-like serine proteases. [4,5] Further representative examples of ligands known to be suitable for affinity chromatography protocols include the 27-mer peptide derived from the exon 1B of the endogenous calpain inhibitor calpastatin, [6] the propeptide of cathepsin D, [7] and the peptide boronic acid AlaAla-boroVal directed against human neutrophil elastase. [8] Human tryptases represent a group of serine proteases (clan PA(S), family S1) with trypsin-like activity that are almost exclusively expressed in mast cells.…”

Effect of the Microenvironment on the Mode of Action of Immobilized Enzymes

“…More recently several enzymes have been bound, via their amino groups, to cellulose activated by sym-trichlorotriazine (cyanuric chloride) (49)(50)(51) or by B dichloroqm-triazinyl dyestuff (Procion brilliant orange MGS) (52, 53), as well as to Sephadex or Sepharose, activated by cyanogen bromide (54)(55)(56)(57)(58). A polymeric acylating reagent, ethylene-maleic anhydride (1 : 1) copolymer (EMA), has been successfully used for the preparation of polyanionic water-insoluble derivatives of enzymes, antigens, and protein enzyme inhibitors (10,(59)(60)(61)(62)(63)(64)(65)(66). EMA-papain end EMAchymotrypsin conjugates have been converted into polycationic or polyalcohol derivatives by coupling the polycarboxylic enzyme derivative with N,N-dimethylaminopropylamine or propanolamine, respectively, in the presence of carbodiimide (67).…”