MDA231_3BrP__flux_HK_GAPDH

Selected Cell

Cell:

Value:

Metadata Organism_Sample Data_protein Data_enzyme activity Data_flux Result_MDA231_3BrP

Metadata	Values (examples)

Asset Title	MDA231_3-Bromopyruvate titration in intact cells: Enzyme and flux inhibition data
Uploader	Theresa Kouril
Uploader SEEK ID	261
Project	Project	Glucose metabolism in cancer cells

ASSAY
Assay SEEK ID
Assay Title	Enzyme and flux inhibitiom in intact cells
Assay_type	enzymaticAssay	extracellular metabolite concentration	protein concentration
Technology_type	plate reader based assays	plate reader based assays	Bradford
Description
Experimentalist	Theresa Kouril
Date
SOP
Publication (optional)

Experimental_conditions
Item	temperature	gas_flow	growth_medium
Compound (if concentration)		CO2	DMEM
Unit	°C	%
Start_value (optional)	37	5	low glucose
End_value (optional)
Comments

Culture growth	Batch

FACTORS_STUDIED
Item	FactorsStudied	FactorsStudied
Compound (if concentration)	3-Bromopyruvate
Unit	mM
Start_value (optional)	0
End_value (optional)	0.75
SD (optional)
Comments

Sample Name			chatacteristics[Treatments (Factors Studied)]	experimental details	comment [replica type]	comment[date_experiment]
	organism	chatacteristics [cell line]	specific_concentration	volume		date
1	Homo sapiens	MDA231	0-0.75mM 3BrP	7ml LG DMEM, 100 mm dishes	biological	2020-12-31T0:0:0+0000
2	Homo sapiens	MDA231	0-0.75mM 3BrP	7ml LG DMEM, 100 mm dishes	biological	2020-12-10T0:0:0+0000
3	Homo sapiens	MDA231	0-0.75mM 3BrP	7ml LG DMEM, 100 mm dishes	biological	2020-12-10T0:0:0+0000
4	Homo sapiens	MDA231	0-0.3mM 3BrP	10 ml LG DMEM, 100 mm dishes	biological	2021-01-21T0:0:0+0000
5	Homo sapiens	MDA231	0-0.3mM 3BrP	10 ml LG DMEM, 100 mm dishes	biological	2021-01-21T0:0:0+0000
6	Homo sapiens	MDA231	0-0.3mM 3BrP	10 ml LG DMEM, 100 mm dishes	biological	2021-01-21T0:0:0+0000
7	Homo sapiens	MDA231	0-0.2mM 3BrP	7ml LG DMEM, 100 mm dishes	biological	2021-01-27T0:0:0+0000
8	Homo sapiens	MDA231	0-0.2mM 3BrP	7ml LG DMEM, 100 mm dishes	biological	2021-01-27T0:0:0+0000




Note: this sheet allows you to describe your samples in as much detail as required. Sample names can be used in the data matrix file to relate different conditions to different experimental measurements

Brdaford assay
Protein concentration	[3BrP]	sample	sample	sample
		1	2	3
	(μM)	mg/ml	mg/ml	mg/ml
	0	1.1	2.4	2.1
	150	1.2	2.6	2.2
	300	1.3	2.4	2.3
	450	1.2	2.3	2
	600	0.9	2.3	1.9
	750	0.6	1	0.4		As refelcted in the protein concentration, cells start to lift during the 8h period of the experiment and were therefore not consindered in the analysis

	[3BrP]	sample	sample	sample	sample	sample
		4	5	6	7	8
Protein concentration	(μM)	mg/ml	mg/ml	mg/ml	mg/ml	mg/ml
	0	1.9	1.9	1.9	1.4	1.6
	100	1.9	1.9	2.1	1.7	1.7
	200	2	1.7	2.3	1.7	1.7
	300	2	2	2.1

Enzymatic Assay
Enzyme	[3BrP]	sample	sample	sample	sample	sample	sample	sample	sample
		1	2	3	4	5	6	7	8
	(μM)	activity (nmol/min/mg)	activity (nmol/min/mg)	activity (nmol/min/mg)	activity (nmol/min/mg)	activity (nmol/min/mg)	activity (nmol/min/mg)	activity (nmol/min/mg)	activity (nmol/min/mg)
Hexokinase	0	72	109	112	78	79	98	89	84
	100	76	76	75	80	82
	150	70	98	106
	200	67	73	69	76	69
	300	60	89	98	57	65	67
	450	58	86	84
	600	55	72	77
	750	30	47	66
GAPDH	0	445	558	646	496	438	416	668	584
	100	324	296	320	436	397
	150	235	383	376
	200	165	166	195	296	256
	300	158	201	246	102	129	133
	450	119	152	210
	600	126	109	127
	750	114	105	147

Enzyme	[3BrP]	sample	sample	sample	sample	sample
		4	5	6	7	8
	(μM)	activity (nmol/min/mg)	activity (nmol/min/mg)	activity (nmol/min/mg)	activity (nmol/min/mg)	activity (nmol/min/mg)
Hexokinase	0	78	79	98	89	84
	100	76	76	75	80	82
	200	67	73	69	76	69
	300	57	65	67
GAPDH	0	496	438	416	668	584
	100	324	296	320	436	397
	200	165	166	195	296	256
	300	102	129	133

Flux
Metabolite	[3BrP]	sample	sample	sample	sample	sample	sample	sample	sample
		1	2	3	4	5	6	7	8
	(μM)	J (nmol/min/mg)	J (nmol/min/mg)	J (nmol/min/mg)	J (nmol/min/mg)	J (nmol/min/mg)	J (nmol/min/mg)	J (nmol/min/mg)	J (nmol/min/mg)
Lacate	0	89	99	118	91	83	86	98	94
	100	88	91	83	92	91
	150	98	86	121
	200	84	89	78	86	85
	300	78	82	94	61	81	74
	450	65	64	84
	600	63	43	42
	750	32	35	55
Glucose	0	36	52	56	42	37	33	57	48
	100	44	34	30	46	50
	150	40	46	57
	200	38	24	26	42	43
	300	25	41	41	35	37	24
	450	19	32	30
	600	20	17	19
	750	2	19	20

Metabolite	[3BrP]	sample	sample	sample	sample	sample
		4	5	6	7	8
	(μM)	J (nmol/min/mg)	J (nmol/min/mg)	J (nmol/min/mg)	J (nmol/min/mg)	J (nmol/min/mg)
Lactate	0	91	83	86	98	94
	100	88	91	83	92	91
	200	84	89	78	86	85
	300	61	81	74
Glucose	0	42	37	33	57	48
	100	44	34	30	46	50
	200	38	24	26	42	43
	300	35	37	24

Sample	[3BrP] (μM)	Relative Flux		Relative Activity
		Lactate	Glucose	HK	GAPDH
1	0	1.0001	1.00001	1.0001	1.00001
	150	0.9	0.9	0.9722222222222222	0.5280898876404494
	300	0.8764044943820225	0.6944444444444444	0.8333333333333334	0.3550561797752809
	450	0.7303370786516854	0.5277777777777778	0.8055555555555556	0.26741573033707866
	600	0.7078651685393258	0.5555555555555556	0.7638888888888888	0.28314606741573034
2	0	1.000001	1	1	1
	150	0.8686868686868687	0.8846153846153846	0.8990825688073395	0.6863799283154122
	300	0.8282828282828283	0.7884615384615384	0.8165137614678899	0.3602150537634409
	450	0.6464646464646465	0.6153846153846154	0.7889908256880734	0.2724014336917563
	600	0.43434343434343436	0.3269230769230769	0.6605504587155964	0.1953405017921147
3	0	1	1	1	1
	150	1.0254237288135593	1.0178571428571428	0.9464285714285714	0.5820433436532507
	300	0.7966101694915254	0.7321428571428571	0.875	0.38080495356037153
	450	0.711864406779661	0.5357142857142857	0.75	0.32507739938080493
	600	0.3559322033898305	0.3392857142857143	0.6875	0.19659442724458204
4	0	1.01	1.01	1.01	1.001
	100	0.967032967032967	1.0476190476190477	0.9743589743589743	0.6532258064516129
	200	0.9230769230769231	0.9047619047619048	0.8589743589743589	0.3326612903225806
	300	0.6703296703296703	0.8333333333333334	0.7307692307692307	0.2056451612903226
5	0	1	1	1	1
	100	1.0963855421686748	0.918918918918919	0.9620253164556962	0.6757990867579908
	200	1.072289156626506	0.6486486486486487	0.9240506329113924	0.3789954337899543
	300	0.9759036144578314	1	0.8227848101265823	0.2945205479452055
6	0	1	1	1	1
	100	0.9651162790697675	0.9090909090909091	0.7653061224489796	0.7692307692307693
	200	0.9069767441860465	0.7878787878787878	0.7040816326530612	0.46875
	300	0.8604651162790697	0.7272727272727273	0.6836734693877551	0.31971153846153844
7	0	1.0001	1.001	1.001	1.001
	100	0.9387755102040817	0.8070175438596491	0.898876404494382	0.6526946107784432
	200	0.8775510204081632	0.7368421052631579	0.8539325842696629	0.4431137724550898
8	0	1.00001	1.00001	1	1
	100	0.9680851063829787	1.0416666666666667	0.9761904761904762	0.6797945205479452
	200	0.9042553191489362	0.8958333333333334	0.8214285714285714	0.4383561643835616