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Using cytochome c to monitor electron transport and inhibition in beef heart submitochondrial particles *
Author(s) -
Melin Amanda D.,
LohmeierVogel Elke M.
Publication year - 2004
Publication title -
biochemistry and molecular biology education
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.34
H-Index - 39
eISSN - 1539-3429
pISSN - 1470-8175
DOI - 10.1002/bmb.2004.494032010306
Subject(s) - submitochondrial particle , redox , electron transport chain , cytochrome c , cytochrome , respiratory chain , chemistry , antimycin a , rotenone , coenzyme q – cytochrome c reductase , biophysics , biochemistry , mitochondrion , enzyme , organic chemistry , biology
We present a two‐part undergraduate laboratory exercise. In the first part, electron transport in bovine heart submitochondrial particles causing reduction of cytochrome c is monitored at 550 nm. Redox‐active dyes have historically been used in most previous undergraduate laboratory exercises of this sort but do not demonstrate respiratory inhibition by antimycin A and rotenone. By using cytochrome c instead of redox‐active dyes, it is possible to observe inhibition of electron transport in the presence of the aforementioned respiratory inhibitors. In the second part, students are asked to design a soluble redox chain between NADH and cytochrome c using catalytic amounts of redox‐active dyes. The students are also responsible for designing the assays and control. The entire experiment can be performed in 3 h with single‐beam spectrophotometers that are currently used in most undergraduate teaching laboratories. This exercise is suitable for large undergraduate classes of over 200 students and can be performed either by a single student or a student pair.