Biliverdin reductase A activity

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Focus: BVRA is an enzyme that reduces biliverdin to bilirubin. NADPH is consumed in the reaction.

Draft scope: There are some open questions in the right method for measuring BVRA activity. The question about blank activity will be discussed.

Variables:

  • Total activity. Definition: Observed increase at A460 per minute in reaction mix consisting of buffer, biliverdin, cytosol and NADPH.
  • Blank. Definition: Observed increase at A460nm per minute in reaction mix consisting of buffer, biliverdin and cytosol.
  • Putatively specific avtivity. Definition: Total activity - blank.

Statement: Total activity is a better measure of actual BVRA activity than putatively specific activity.

Argumentation:

← Energy sources (NADPH) are available in the cytosol.

← Total activity is a specific measure of BVRA

← Activity=0 if no cytosol (own observation)
→ When no cytosol, there is no BVRA
← Activity=0 if no BV (own observation)
← It is unlikely that the reaction is unspecific
← Blank is large compared with total