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Expression, Purification, Characterization, and X-Ray Analysis of Selenomethionine 215 Variant of Leukocyte Collagenase (1997)

Pieper, Michael ; Betz, Michael ; Budisa, Nediljko ; [et al.]

Springer

The protein journal 16 (1997), S. 637-650

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ISSN: 1573-4943

Keywords: Matrix metalloproteinases ; Met-turn ; Selenomethionine ; conformational stability ; X-ray crystallography

Source: Springer Online Journal Archives 1860-2000

Topics: Chemistry and Pharmacology

Notes: Abstract Matrix metalloproteinases belong to the superfamily of metzincins containing, besides a similar topology and a strictly conserved zinc environment, a 1,4-tight turn with a strictly conserved methionine residue at position three (the so called Met-turn [Bode et al. (1993) FEBS 331, 134–140; Stöcker et al. (1995) Protein Sci. 4, 823–840]. The distal S–CH3 moiety of this methionine residue forms the hydrophobic basement of the three His residues liganding the catalytic zinc ion. To assess the importance of this methionine, we have expressed the catalytic domain of neutrophil collagenase (rHNC, residues Met80–Gly242) in the methionine auxotrophic Escherichia coli strain B834[DE3](hsd metB), with the two methionine residues replaced by Selenomethionine. Complete replacement was confirmed by amino acid analysis and electrospray mass spectrometry. The folded and purified enzyme retained its catalytic activity, but showed modifications which are reflected in changed kinetic parameters. The Met215SeMet substitution caused a decrease in conformational stability upon urea denaturation. The X-ray crystal structure of this Selenomethionine rHNC was virtually identical to that of the wild-type catalytic domain except for a very faint local disturbance around the sulfur-seleno substitution site.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1023/A:1026327125333

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Diels-Alder-Reaktionen von o-Benzochinonen mit Fulvenen (1978)

Friedrichsen, Willy ; Betz, Michael ; Büldt, Eggert ; [et al.]

Weinheim : Wiley-Blackwell

Liebigs Annalen 1978 (1978), S. 440-472

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ISSN: 0075-4617

Keywords: Chemistry ; Organic Chemistry

Source: Wiley InterScience Backfile Collection 1832-2000

Topics: Chemistry and Pharmacology

Description / Table of Contents: Diels-Alder Reactions of o-Benzoquinones with Fulveneso-Benzoquinone (1a) and alkyl-substituted o-benzoquinones 1b-d react with fulvenes 2a-d in a stereoselective and regiospecific cycloaddition to give endo-dicarbonyl compounds 3a-o and 4a, b. Photodecarbonylation and dehydrogenation of these adducts yields benzofulvenes 6, which could be synthesized from indanones 8 by an independent route.

Notes: o-Benzochinon (1a) und alkylsubstituierte o-Benzochinone 1b-d reagieren mit Fulvenen 2a-d in einer stereoselektiv und regiospezifisch verlaufenden Cycloaddition unter Bildung von endo-Dicarbonylverbindungen 3a-o und 4a, b. Aus diesen sind durch Photodecarbonylierung und Dehydrierung Benzofulvene 6 zugänglich, die auch auf unabhängigem Wege aus Indanonen 8 dargestellt wurden.

Additional Material: 12 Tab.