Cookies help us deliver our services. By using our services, you agree to our use of cookies. More information

Difference between revisions of "ATP"

From Bioblast
Line 1: Line 1:
{{MitoPedia
{{MitoPedia
|abbr=T
|abbr=T
|description='''Adenosine triphosphate''' is a nucleotid and functions as the major carrier o fchemical energy in the cells. As it transfers its energy to other molecules, it looses its terminal phosphate group and becomes adenosine diphosphate [[(ADP)]] .
|description='''Adenosine triphosphate''' is a nucleotid and functions as the major carrier o fchemical energy in the cells. As it transfers its energy to other molecules, it looses its terminal phosphate group and becomes adenosine diphosphate ([[ADP]]) .
|info=[[MiPNet03.02]]; [[MiPNet09.12]]
|info=[[MiPNet03.02]]; [[MiPNet09.12]]
|type=Substrate PS
|type=Substrate PS

Revision as of 16:06, 21 June 2012


high-resolution terminology - matching measurements at high-resolution


ATP

Description

Adenosine triphosphate is a nucleotid and functions as the major carrier o fchemical energy in the cells. As it transfers its energy to other molecules, it looses its terminal phosphate group and becomes adenosine diphosphate (ADP) .

Abbreviation: T

Reference: MiPNet03.02; MiPNet09.12


MitoPedia topics: Substrate and metabolite 


Application in HRR

T: ATP (Adenosine 5'-triphosphate disodium salt hydrate, C10H14N5O13P3Na2* (H2O)x); Sigma A 2383, 5 g, store at R.T.; FW = 614.1 containing 3.5 mol H2O/mol (FWanhydrous basis = 551.1). Alternative source: Merck 1191-1GM (Calbiochem, Adenosine 5′-Triphosphate Disodium Salt; FW = 551.2; store at -20 °C).


Caution: Chemicals stored in the fridge or freezer should be allowed to reach room temperature before opening.


Preparation of 500 mM ATP stock solution (dissolved in H2O):

1) weigh 614.1 mg of ATP (FW = 614.1 containing 3.5 mol H2O/mol)
2) add 1.2 ml H2O
3) neutralize with 5 N KOH (approx. 400 µl) - ATP will dissolve after addition of KOH! (this is Solution A)
4) check pH and adjust to 7 if necessary
5) adjust final volume to 2 ml and divide into 0.2 ml portions
6) store frozen at -80 °C


Preparation of 500 mM ATP stock solution with 3 mM free Mg2+:

To keep free [MgMg2+] constant during respiration measurement mix ATP with MgCl2 (0.8 mol MgCl2 /mol ATP).

MgCl2 (Scharlau MA0036: MgCl2.6H2O, FW = 203.3)

1) add 162.64 mg MgCl2 to Solution A! White precipitate will occur, which will be dissolved after 1-2 min of stirring on magnetic stirrer at room temperature!
2) follow steps 4-6 of the above instructions
Comment: In some cases it was observed, that after defreezing of the 'ATP-Mg' a white precipitate has formed again. If this is the case we recommend to prepare the 500 mM ATP stock solution and a MgCl2 solution (0.8 mol MgCl2 /mol ATP) seperately and perform a titration of both solutions in immediate succession.


Oxygraph-2k Manual Titrations: MiPNet09.12

  • Titration volume: 4-20 µl using a 25 µl syringe
  • Final conc. in 2 ml O2k-chamber: 1-5 mM