Using cytochome c to monitor electron transport and inhibition in beef heart submitochondrial particles

• Published in: Biochemistry and molecular biology education Vol. 32; no. 1; pp. 39 - 44
• Main Authors: Melin, Amanda D., Lohmeier‐Vogel, Elke M.
• Format: Journal Article
• Language: English
• Published: USA John Wiley & Sons Inc 01-01-2004; John Wiley & Sons, Inc
• Subjects: Animals; Chemistry; cytochrome c; electron transport; Inhibition; Metabolism; Science Experiments; Science Instruction; Science Laboratories; Submitochondrial particles; Teaching Methods; Undergraduate Students
• ISSN: 1470-8175; 1539-3429
• DOI: 10.1002/bmb.2004.494032010306

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.