Question

2. Based on your experimental observations, what can you conclude concerning the distribution of succinate dehydrogenase? Is the distribution what you would have expected? Explain why or why not.

Answer 1

Ans Succinate clehydkoge nase (SDM) i s an en3yme found in the inner mito chondrial mem brone, w when studying the cihic acid cycle hrch makes it an easy tuhget tho isolate e naymafe octvity ill be olectem ned by mitochondvrial ocfemafion Centrifugation Methed n ikalah mitchandvia, we vomoved with a scalpel tany iedefe hen calliHaver Pom the outer 2-3 mm swpace Then,--weyhindedーHe t ssue with a pestle in a mortar n 40 ml f cl-cole mannitol qpunding buller for 4 min Now, wegltered the Suspensionな1s9&eze(5th solution out- througt Jour layers of' cheesecloth int hree chilled is m? Cent hueahom tube. min, and de cantld the supernataut iuto a chilled 5o ml cents tube ter we ye-spun the pitrodc solutom ut to,aro x qma or 30 mint 0-4ic and disc ondedl the Supe natunt U in th smk leaving the pellet Then, we added 7.o ml ot ice coled mannitol assay bufes to the mihu chendiel pellet inel seraped andl mixed the mto chan dki pellet from the vwall of centrijuige tube with spatula Re sult The spectvo pho tomet teble er sesults we obtained awre presented in he table The qpuatest abgorbance reading toas oktuined test tube 4 at 363 abs, which is because malonadehe Competitos nhibioy, is present almg the pubs iete succinate So even houglh more mtochondria may hove been pre set in Tube 8 te they were not free to imte sot with the ohe in soluthon. A Becond pu asen may be thf ho much DaIP wos addled to Tube & so here asn excess DCIP uagent in hot samp