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Investigation Of The Respiratory Chain In Mitochndria. The experiments were carried out to investigate electron transport of the respiratory chain of yeast mitochondria and to examine the effects of different compounds on the chain. This was done using s

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Introduction

Introduction The experiments were carried out to investigate electron transport of the respiratory chain of yeast mitochondria and to examine the effects of different compounds on the chain. This was done using spectrometry and the absorbance spectral properties of a redox dye (DCPIP), which acts as an artificial electron donor/acceptor. Method For safety guidelines, the apparatus and the techniques employed when carrying out the experiment, see the attached, workshop handout. Results Results Table Experiment A Minutes Experiment 1 (Part 2) Absorbance Experiment 1 (Part 3) Absorbance Experiment 1 (Part 4) Absorbance Experiment 1(Part 5) Absorbance 0 0.96 0.95 0.91 0.54 0.5 0.96 0.95 0.895 0.54 1 0.96 0.95 0.87 0.54 1.5 0.96 0.94 0.85 0.54 2 0.96 0.94 0.83 0.54 2.5 0.96 0.94 0.81 0.54 3 0.96 0.94 0.79 0.545 3.5 0.96 0.94 0.77 0.545 4 0.96 0.935 0.76 0.545 4.5 0.96 0.935 0.735 0.545 5 0.96 0.935 0.72 0.55 5.5 0.705 6 0.69 6.5 0.655 7 0.65 7.5 0.64 8 0.625 8.5 0.61 9 0.6 9.5 0.585 10 0.57 10.5 0.56 11 0.55 11.5 0.54 12 0.53 12.5 0.515 13 0.51 13.5 0.49 14 0.48 14.5 0.475 15 0.46 Results Table Experiment B Minutes Experiment B (Part 1) ...read more.

Middle

Experiment B Maximum rate of DCPIP reduction calculations: Part 1 = 0.91-0.40 / 263 = 0.51 / 263 x 1000 = 1.939163498 nmol min-1 Part 2 = 0.84-0.75 / 263 = 0.09 / 263 x 1000 = 0.342205323 nmol min-1 Part 3 = 0.75-0.495 / 263 = 0.255 / 263 x 1000 = 0.969581749 nmol min-1 Discussion The chain complex which donates electrons to DCPIP is Chain III. This is because when cyanide is added, the DCPIP becomes gradually colourless as can be seen by the spectrometer readings (dropping from 0.19 to 0.46) over the 15 minute run. Because DCPIP loses its colour, this means that it is still being reduced and if complex 4 was the donator then this would have stopped when the cyanide was added. When the Antimycin A was added the reduction stopped which meant that electrons were no longer being donated. Because the reduction stopped when Complex III was inhibited, it can be concluded that Complex III is the electron donor for DCPIP. ...read more.

Conclusion

This is because when there was only cyanide in the mitochondrial assay and therefore, Complex IV was inhibited, the reduction of DCPIP was at a fairly constant rate and also reached it highest rate of reduction over 15 minutes as shown by the gradient of the line on the plot.(part 1 line on plot). When compound x was added the reduction of DCPIP was reduced. The mode of action of compound x could either be a cyanide inhibitor, which would slow the inhibition of Complex IV also and so would allow DCPIP to donate electrons again, slowing the reduction of DCPIP. It could also be another redox agent, which by also accepting electrons, would lower the reduction of DCPIP. I have a high confidence in the reliability of the results. From what we know respiratory inhibitors, it is clear that the experiment acted as it was expected to. However, the exception is in the results from the assay which contained compound x. This is because as an unknown compound, it can not be said that this experiment acted as it should, as what effect compound x has is unknown. ...read more.

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