Lab 3

Question 1

Differential centrifugation is used to seperate cellular components- At 200rpm or 600xg pellet=—-debirs=—-

Answer 1

pellet=whole cells
debris=nuclei and cell debris

Question 2

Differential centrifugation is used to seperate cellular components- At 10,000rpm or 1500xg
pellet=

Answer 2

pellet=mitochondria, cell membrane

Question 3

Differential centrifugation is used to seperate cellular components- At 40,000rpm or 100,000xg

pellet=

Answer 3

ribosomes

Question 4

As centrifugal force is increased ,—-and —-dense componets are isolated

Answer 4

smaller and less dense compnents are isolated

Question 5

—-are transferred through compex I-IV to terminal electron acceptor O2

Answer 5

Electrons

Question 6

Ubiquinone-

Answer 6

transfers electrons from Complex I and II to bComplex III

Question 7

Cytochrome C-

Answer 7

transferes eletrons from complex III to Complex IV

Question 8

—-reduces oxygen to water

Answer 8

Complex IV

Question 9

Succinate dehydrogenase(SDH)

Answer 9

oxidizes succinate to fumarate in Compex 2, releasing two eletrons into ETC

-this can be assessed by a ETC inhibitor and electron acceptor

Question 10

—-is a downstream ETC inhibitor

Answer 10

Sodiumn azide

Question 11

sodium azide blocks eletron transfer by

Answer 11

cytochrone c and ubiquinone

Question 12

How can we specifically assess SDH activity not just complex I and III activity?

Answer 12

use sodium malonate to inhibit SDH and compare

Question 13

—-is an SDH inhibitor

Answer 13

sodium malonate

Question 14

Sodium malonate is —-of succinate letting it competivitly —-SDH by—-. This can induce a —. This can aslo increase production of —and free—

Answer 14

a structural analolg of succinate
competivilty inhibits
induce a mitochondrial potnetial collapse
production of reactive oxygen species and free eletrons

Question 15

Excess electrons accepted by an artifical electron acceptor-

Answer 15

2,6 dichlorophenol indophenol (DCIP)
this is a indirect measure of enzyme activity

Question 16

Summarize procedure

Answer 16

isolate mitochondria by centrifugation
setup enzyme reactions with-succinate, sodium azide, malonate, DCIP inot MF and MFF

Question 17

How to calculate RCF

Answer 17

(1.119x10-5)(rpm)2(r)

Question 18

Difference between a gravity to deliever and blowout

Answer 18

Blowout is the appropriate amount is delievered by the last blow

gravity there will be fluid left but the appropriate amount

Question 19

When using a Peleus bulb explain. the valves A S E

Answer 19

A=expel the air from buld while holding this down
S=suction up the liquid
E=empty the fluid