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. 1972 Dec;130(3):825–832. doi: 10.1042/bj1300825

The ionogenic nature of the secretory-granule membrane. Electrokinetic properties of isolated chromaffin granules

E K Matthews 1, R J Evans 1, P M Dean 1
PMCID: PMC1174522  PMID: 4664936

Abstract

1. Chromaffin granules isolated from the bovine adrenal medulla possess an electrophoretic mobility of −1.12μm·s−1·cm·V−1, corresponding to a surface ζ potential of −14.4mV and surface charge density of 1.38×10−6C·cm−2. 2. The mobility of chromaffin granules is pH-dependent, indicating an amphoteric surface with an isoelectric point at pH3.0 and acidic groups with a pKa of 3.11. 3. Addition of bi- and ter-valent cations decreased the mobility of chromaffin granules in a dose-dependent fashion with a relative potency of La3+»Mn2+>Ca2+ >Sr2+>Mg2+>Ba2+. 4. Treatment with neuraminidase decreased the mobility of erythrocytes by 84%, whereas chromaffin-granule mobility was decreased by only 14%. This correlates well with the small complement of neuraminic acid present in the granule membrane. 5. The nature, origin and significance of the anionic surface charge of the chromaffin granule is discussed. It is concluded that the net negative charge at the surface of shear derives chiefly from a single type of chemical group, namely −CO2−, contributed by the α-carboxyl group of constituent proteins, the phospholipid phosphatidylserine and, to a lesser extent, the sialic acid component of glycoproteins.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BANGHAM A. D., HEARD D. H., FLEMANS R., SEAMAN G. V. An apparatus for microelectrophoresis of small particles. Nature. 1958 Sep 6;182(4636):642–644. doi: 10.1038/182642a0. [DOI] [PubMed] [Google Scholar]
  2. BERTLER A., CARLSSON A., ROSENGREN E. A method for the fluorimetric determination of adrenaline and noradrenaline in tissues. Acta Physiol Scand. 1958 Dec 15;44(3-4):273–292. doi: 10.1111/j.1748-1716.1958.tb01627.x. [DOI] [PubMed] [Google Scholar]
  3. BLASCHKO H., WELCH A. D. Localization of adrenaline in cytoplasmic particles of the bovine adrenal medulla. Naunyn Schmiedebergs Arch Exp Pathol Pharmakol. 1953;219(1-2):17–22. doi: 10.1007/BF00246245. [DOI] [PubMed] [Google Scholar]
  4. Blaschko H., Firemark H., Smith A. D., Winkler H. Lipids of the adrenal medulla. Lysolecithin, a characteristic constituent of chromaffin granules. Biochem J. 1967 Aug;104(2):545–549. doi: 10.1042/bj1040545. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. COOK G. M., HEARD D. H., SEAMAN G. V. Sialic acids and the electrokinetic charge of the human erythrocyte. Nature. 1961 Jul 1;191:44–47. doi: 10.1038/191044a0. [DOI] [PubMed] [Google Scholar]
  6. DODGE J. T., MITCHELL C., HANAHAN D. J. The preparation and chemical characteristics of hemoglobin-free ghosts of human erythrocytes. Arch Biochem Biophys. 1963 Jan;100:119–130. doi: 10.1016/0003-9861(63)90042-0. [DOI] [PubMed] [Google Scholar]
  7. Douglas W. W. Stimulus-secretion coupling: the concept and clues from chromaffin and other cells. Br J Pharmacol. 1968 Nov;34(3):451–474. doi: 10.1111/j.1476-5381.1968.tb08474.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. EYLAR E. H., MADOFF M. A., BRODY O. V., ONCLEY J. L. The contribution of sialic acid to the surface charge of the erythrocyte. J Biol Chem. 1962 Jun;237:1992–2000. [PubMed] [Google Scholar]
  9. HAGGENDAL J. Fluorimetric determination of 3-O-methylated derivatives of adrenaline and noradrenaline in tissues and body fluids. Acta Physiol Scand. 1962 Nov-Dec;56:258–266. doi: 10.1111/j.1748-1716.1962.tb02503.x. [DOI] [PubMed] [Google Scholar]
  10. HILLARP N. A., LAGERSTEDT S., NILSON B. The isolation of a granular fraction from the suprarenal medulla, containing the sympathomimetic catechol amines. Acta Physiol Scand. 1953 Oct 6;29(2-3):251–263. doi: 10.1111/j.1748-1716.1953.tb01022.x. [DOI] [PubMed] [Google Scholar]
  11. HODGKIN A. L., KEYNES R. D. Movements of labelled calcium in squid giant axons. J Physiol. 1957 Sep 30;138(2):253–281. doi: 10.1113/jphysiol.1957.sp005850. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. IVERSEN L. L. THE UPTAKE OF NORADRENALINE BY THE ISOLATED PERFUSED RAT HEART. Br J Pharmacol Chemother. 1963 Dec;21:523–537. doi: 10.1111/j.1476-5381.1963.tb02020.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Jarrott B. Occurrence and properties of monoamine oxidase in adrenergic neurons. J Neurochem. 1971 Jan;18(1):7–16. doi: 10.1111/j.1471-4159.1971.tb00162.x. [DOI] [PubMed] [Google Scholar]
  14. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  15. Poisner A. M., Trifaró J. M. The role of ATP and ATPase in the release of catecholamines from the adrenal medulla. I. ATP-evoked release of catecholamines, ATP, and protein from isolated chromaffin granules. Mol Pharmacol. 1967 Nov;3(6):561–571. [PubMed] [Google Scholar]
  16. Smith A. D., Winkler H. Purification and properties of an acidic protein from chromaffin granules of bovine adrenal medulla. Biochem J. 1967 May;103(2):483–492. doi: 10.1042/bj1030483. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Tenforde T. Microelectrophoretic studies on the surface chemistry of erythrocytes. Adv Biol Med Phys. 1970;13:43–105. doi: 10.1016/b978-0-12-005213-4.50006-5. [DOI] [PubMed] [Google Scholar]
  18. WARREN L. The thiobarbituric acid assay of sialic acids. J Biol Chem. 1959 Aug;234(8):1971–1975. [PubMed] [Google Scholar]
  19. Winkler H., Hörtnagl H., Smith A. D. Membranes of the adrenal medulla. Behaviour of insoluble proteins of chromaffin granules on gel electrophoresis. Biochem J. 1970 Jun;118(2):303–310. doi: 10.1042/bj1180303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. von EULER U., LISHAJKO F. Improved technique for the fluorimetric estimation of catecholamines. Acta Physiol Scand. 1961 Apr;51:348–355. doi: 10.1111/j.1748-1716.1961.tb02128.x. [DOI] [PubMed] [Google Scholar]

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