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Non-protein-coding regions There are nearly 25% of mtDNA mutations occurring in non-protein-coding regions including tRNAs and the highly conserved ribosomal RNA [125]

Also valine has been proven important for the oxidation of the reduced flavin by molecular oxygen in choline oxidase and it has been reported that replacement of Val464 with alanine in the enzyme results in a twofold decrease in the limiting rate constant for flavin reduction (k red ) and less than fivefold decrease in the equilibrium constant for formation of the enzymesubstrate complex (K d ) 52

For example, Gao et al

Membranes were blocked for 1 h at room temperature in tris buffered saline with tween 20 (TBST) and 5% non-fat dry milk followed by three washes (15 min for each wash) in TBST with mild shaking