Lipids

, Volume 28, Issue 10, pp 877–882 | Cite as

pH-dissociation characteristics of cardiolipin and its 2′-deoxy analogue

  • Morris Kates
  • Jing-Yi Syz
  • David Gosser
  • Thomas H. Haines
Article

Abstract

Cardiolipin (CL) is found in inner mitochondrial membranes and the plasma membrane of aerobic prokaryotes. CL is tightly bound to those transmembrane enzymes associated with oxidative phosphorylation. CL has earlier been reported to have a single pK at low pH. We have titrated CL in aqueous suspension (bilayers) and in solution in methanol/water (1∶1, vol/vol) and found it to display two different pK values, pK1 at 2.8 and pK2 initially at 7.5 but shifting upwards to 9.5 as the titration proceeds. The unusually high pK2 might be explained by the formation of a unique hydrogen bond in which the free hydroxyl on the central glycerol forms a cyclic intramolecular hydrogen-bonded structure with one protonated phosphate (P-OH group). We have therefore chemically synthesized the 2′-deoxycardiolipin analogue, which lacks the central free hydroxyl group, and measured its pH-dissociation behavior by potentiometric titration, under the same conditions as those for CL. The absence of the hydroxyl group changes the titration dramatically so that the deoxy analogue displays two closely spaced low pK values (pK1=1.8; pK2=4.0). The anomalous titration behavior of the second dissociation constant of CL may be attributed to the participation of the central glycerol OH group in stabilizing the formation of a cyclic hydrogen-bonded monoprotonated form of CL, which may function as a reservoir of protons at relatively high pH. This function may have an important bearing on proton pumping in biological membranes.

Abbreviations

BH

beef heart

CL

cardiolipin (diphosphatidylglycerol)

CL-BH

cardiolipin from beef heart

18∶0-CL (H2-CL-BH)

hydrogenated beef heart cardiolipin

H2-CL-EC

hydrogenated cardiolipin fromE. coli

cy-17∶0 andcy-19∶0

9,10-methylenehexadecanoic and octadecanoic acid, respectively

DAG

diacylglycerols

dCL

2′-deoxycardiolipin (diphosphatidyl-1,3-propanediol)

16∶0-dCL

synthetic deoxycardiolipin with palmitoyl (16∶0) as acyl groups

DPG

dipalmitoylglycerol

DPPA

dipalmitoyl phosphatidic acid

FAB-MS

fast-atom bombardment mass spectrometry

FTIR

Fourier transform infrared spectroscopy

GLC

gas-liquid chromatography

NMR

nuclear magnetic resonance

PA

phosphatidic acid

PPD

phosphatidylpropanediol

PPOP

phosphatidylpropanediolphosphate

TLC

thin-layer chromatography

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Copyright information

© American Oil Chemists’ Society 1993

Authors and Affiliations

  • Morris Kates
    • 1
  • Jing-Yi Syz
    • 2
  • David Gosser
    • 2
  • Thomas H. Haines
    • 2
  1. 1.Department of BiochemistryUniversity of OttawaOttawaCanada
  2. 2.Department of ChemistryCity College of the City University of New YorkNew York

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