Evaluation of genes involved in oxidative phosphorylation in yeast by developing a simple and rapid method to measure mitochondrial ATP synthetic activity

Table 1 Relative ATP specific activity of gene deletion mutants related to either mitochondrial ATP synthesis or mitochondrial fatty acid synthesis

Enzyme	Deleted gene	Relative ATP specific activity (%) ^a	Total protein concentration (mg ml ⁻¹ ) ^b
Parental strain (BY4741)	-	100	0.416
NADH dehydrogenase (Complex I)	NDE2	36 ± 4	0.189
NADH dehydrogenase (Complex I)	NDE1	31 ± 20	0.187
Succinate dehydrogenase/Fumarate reductase (Complex II)	SDH1b	44 ± 18	0.189
	EMI5	53 ± 41	0.187
	SDH4	67 ± 33	0.167
	SDH1	33 ± 11	0.181
	SDH2	104 ± 39	0.285
	FRD1	56 ± 31	0.185
	OSM1	61 ± 16	0.207
Electron transferring-flavoprotein dehydrogenase	CIR2	56 ± 9	0.195
Cytochrome c reductase/Cytochrome bc ₁ complex (Complex III)	COR1	38 ± 10	0.179
	RIP1	37 ± 7	0.179
	QCR6	34 ± 21	0.201
	QCR9	22 ± 26	0.191
	QCR10	72 ± 20	0.187
	QCR8	40 ± 9	0.180
	QCR2	n.d.	0.177
	QCR7	n.d.	0.191
	CYT1	n.d.	0.201
	CBP4	15 ± 10	0.173
	FMP25	29 ± 11	0.203
	CBP3	65 ± 25	0.197
	RCF2	35 ± 13	0.200
	CBP6	27 ± 14	0.190
Cytochrome c oxidase (Complex IV)	COX9	44 ± 16	0.187
	COX6	25 ± 6	0.167
	COX5B	37 ± 11	0.191
	COX12	20 ± 9	0.193
	COX8	62 ± 41	0.175
	COX7	n.d.	0.176
	COX5A	30 ± 10	0.182
ATP synthase (Complex V)	ATP1	32 ± 12	0.177
	ATP2	18 ± 5	0.234
	ATP7	52 ± 28	0.179
	ATP14	52 ± 29	0.187
	ATP20	17 ± 8	0.189
	ATP18	40 ± 16	0.168
	ATP4	98 ± 73	0.206
	ATP15	24 ± 16	0.197
	ATP5	38 ± 4	0.187
	INH1	43 ± 14	0.175
	STF1	43 ± 17	0.171
	STF2	53 ± 10	0.190
ADP/ATP translocator	AAC1	59 ± 7	0.178
ADP/ATP translocator	AAC3	53 ± 24	0.208

^aRelative ATP specific activity of the mutant strains was expressed as a percentage of the specific activity of the parental strain.
^bThe protein concentration of the crude mitochondria solution was determined using the Bradford method [25].
All experiments were conducted at least in triplicate. Values are means ± SD.

ISSN: 1475-2859