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Si ‐face stereospecificity at C5 of coenzyme F 420 for F 420 H 2 oxidase from methanogenic Archaea as determined by mass spectrometry
Author(s) -
Seedorf Henning,
Kahnt Jörg,
Pierik Antonio J.,
Thauer Rudolf K.
Publication year - 2005
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/j.1742-4658.2005.04931.x
Subject(s) - stereospecificity , archaea , mass spectrometry , face (sociological concept) , chemistry , cofactor , analytical chemistry (journal) , environmental chemistry , biochemistry , enzyme , catalysis , chromatography , philosophy , gene , linguistics
Coenzyme F 420 is a 5‐deazaflavin. Upon reduction, 1,5 dihydro‐coenzyme F 420 is formed with a prochiral centre at C5. All the coenzyme F 420 ‐dependent enzymes investigated to date have been shown to be Si ‐face stereospecific with respect to C5 of the deazaflavin, despite most F 420 ‐dependent enzymes being unrelated phylogenetically. In this study, we report that the recently discovered F 420 H 2 oxidase from methanogenic Archaea is also Si ‐face stereospecific. The enzyme was found to catalyse the oxidation of (5 S )‐[5‐ 2 H 1 ]F 420 H 2 with O 2 to [5‐ 1 H]F 420 rather than to [5‐ 2 H]F 420 as determined by MALDI‐TOF MS. (5 S )‐[5‐ 2 H 1 ]F 420 H 2 was generated by stereospecific enzymatic reduction of F 420 with (14a‐ 2 H 2 )‐[14a‐ 2 H 2 ] methylenetetrahydromethanopterin.