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. 1971 Nov;125(1):179–187. doi: 10.1042/bj1250179

The interaction of insulin with phospholipids

M C Perry 1,*, W Tampion 1, J A Lucy 1
PMCID: PMC1178038  PMID: 5158903

Abstract

1. A simple two-phase chloroform–aqueous buffer system was used to investigate the interaction of insulin with phospholipids and other amphipathic substances. 2. The distribution of 125I-labelled insulin in this system was determined after incubation at 37°C. Phosphatidic acid, dicetylphosphoric acid and, to a lesser extent, phosphatidylcholine and cetyltrimethylammonium bromide solubilized 125I-labelled insulin in the chloroform phase, indicating the formation of chloroform-soluble insulin–phospholipid or insulin–amphipath complexes. Phosphatidylethanolamine, sphingomyelin, cholesterol, stearylamine and Triton X-100 were without effect. 3. Formation of insulin–phospholipid complex was confirmed by paper chromatography. 4. The two-phase system was adapted to act as a simple functional system with which to investigate possible effects of insulin on the structural and functional properties of phospholipid micelles in chloroform, by using the distribution of [14C]glucose between the two phases as a monitor of phospholipid–insulin interactions. The ability of phospholipids to solubilize [14C]glucose in chloroform increased in the order phosphatidylcholine<sphingomyelin<phosphatidylethanolamine<phosphatidic acid. Insulin decreased the [14C]glucose solubilized by phosphatidylcholine, phosphatidylethanolamine and phosphatidic acid, but not by sphingomyelin. 5. The significance of these results and the molecular requirements for the formation of insulin–phospholipid complexes in chloroform are discussed.

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Selected References

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  1. Arnold J. D., Pak C. Y. Interaction of soluble proteins with protein monolayers. J Am Oil Chem Soc. 1968 Mar;45(3):128–138. doi: 10.1007/BF02915336. [DOI] [PubMed] [Google Scholar]
  2. Baginski E. S., Foà P. P., Zak B. Microdetermination of inorganic phosphate, phospholipids, and total phosphate in biologic materials. Clin Chem. 1967 Apr;13(4):326–332. [PubMed] [Google Scholar]
  3. Bangham A. D., Standish M. M., Watkins J. C. Diffusion of univalent ions across the lamellae of swollen phospholipids. J Mol Biol. 1965 Aug;13(1):238–252. doi: 10.1016/s0022-2836(65)80093-6. [DOI] [PubMed] [Google Scholar]
  4. Blecher M. Phospholipase C and mechanisms of action of insulin and cortisol on glucose entry into free adipose cells. Biochem Biophys Res Commun. 1965 Nov 8;21(3):202–209. doi: 10.1016/0006-291x(65)90272-x. [DOI] [PubMed] [Google Scholar]
  5. Cuatrecasas P. Interaction of insulin with the cell membrane: the primary action of insulin. Proc Natl Acad Sci U S A. 1969 Jun;63(2):450–457. doi: 10.1073/pnas.63.2.450. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. DAS M. L., CRANE F. L. PROTEOLIPIDS. I. FORMATION OF PHOSPHOLIPID-CYTOCHROME C COMPLEXES. Biochemistry. 1964 May;3:696–700. doi: 10.1021/bi00893a017. [DOI] [PubMed] [Google Scholar]
  7. DAS M. L., HAAK E. D., CRANE F. L. PROTEOLIPIDS. IV. FORMATION OF COMPLEXES BETWEEN CYTOCHROME C AND PURIFIED PHOSPHOLIPIDS. Biochemistry. 1965 May;4:859–865. doi: 10.1021/bi00881a010. [DOI] [PubMed] [Google Scholar]
  8. DAS M. L., HIRATSUKA H., MACHINIST J. M., CRANE F. L. The proteolipids of cytochrome c. Biochim Biophys Acta. 1962 Jul 2;60:433–434. doi: 10.1016/0006-3002(62)90427-4. [DOI] [PubMed] [Google Scholar]
  9. Deamer D. W., Cornwell D. G. Calcium action on fatty acid and phospholipid monolayers and its relation to the cell membrane. Biochim Biophys Acta. 1966 Jun 1;116(3):555–562. doi: 10.1016/0005-2760(66)90125-1. [DOI] [PubMed] [Google Scholar]
  10. FENTON E. L. A method for the assay of insulin by paper chromatography. Biochem J. 1959 Mar;71(3):507–513. doi: 10.1042/bj0710507. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. GREEN D. E., FLEISCHER S. THE ROLE OF LIPIDS IN MITOCHONDRIAL ELECTRON TRANSFER AND OXIDATIVE PHOSPHORYLATION. Biochim Biophys Acta. 1963 Oct 22;70:554–582. doi: 10.1016/0006-3002(63)90793-5. [DOI] [PubMed] [Google Scholar]
  12. Green F. A. Binding of phosphatidylcholine-14C to glass. J Lipid Res. 1969 Nov;10(6):710–711. [PubMed] [Google Scholar]
  13. Hanai T., Haydon D. A., Taylor J. The influence of lipid composition and of some adsorbed proteins on the capacitance of black hydrocarbon membranes. J Theor Biol. 1965 Nov;9(3):422–432. doi: 10.1016/0022-5193(65)90041-x. [DOI] [PubMed] [Google Scholar]
  14. Hart C. J., Leslie R. B., Davis M. A., Lawrence G. A. The interaction between cytochrome c and purified phospholipids. Biochim Biophys Acta. 1969;193(2):308–318. doi: 10.1016/0005-2736(69)90191-6. [DOI] [PubMed] [Google Scholar]
  15. Jung C. Y., Chaney J. E., LeFevre P. G. Enhanced migration of glucose from water into chloroform in presence of phospholipids. Arch Biochem Biophys. 1968 Aug;126(2):664–676. doi: 10.1016/0003-9861(68)90454-2. [DOI] [PubMed] [Google Scholar]
  16. Kafka M. S., Pak C. Y. Effects of polypeptide and protein hormones on lipid monolayers. I. Effect of insulin and parathyroid hormone on monomolecular films of monooctadecyl phosphate and stearic acid. J Gen Physiol. 1969 Jul;54(1):134–143. doi: 10.1085/jgp.54.1.134. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kafka M. S., Pak C. Y. The effect of polypeptide hormones on lipid monolayers. ii. The effect of insulin analogues, vasopressin, oxytocin, thyrocalcitonin, adrenocorticotropin, and 3',5'-cyclic AMP on the uptake of Ca2+ by monomolecular films of monooctadecyl phosphate. Biochim Biophys Acta. 1969 Oct 14;193(1):117–123. doi: 10.1016/0005-2736(69)90064-9. [DOI] [PubMed] [Google Scholar]
  18. Kuo J. F. Comparisons of the effects of Bacillus subtilis protease, type VIII (subtilopeptidase A), and insulin on isolated adipose cells. II. Antilipolytic action. J Biol Chem. 1968 Jan 10;243(1):211–215. [PubMed] [Google Scholar]
  19. Kuo J. F., Holmlund C. E., Dill I. K. The effect of proteolytic enzymes on isolated adipose cells. Life Sci. 1966 Dec;5(24):2257–2262. doi: 10.1016/0024-3205(66)90060-9. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. LUCY J. A. GLOBULAR LIPID MICELLES AND CELL MEMBRANES. J Theor Biol. 1964 Sep;7:360–373. doi: 10.1016/0022-5193(64)90080-3. [DOI] [PubMed] [Google Scholar]
  22. Papahadjopoulos D., Miller N. Phospholipid model membranes. I. Structural characteristics of hydrated liquid crystals. Biochim Biophys Acta. 1967 Sep 9;135(4):624–638. doi: 10.1016/0005-2736(67)90094-6. [DOI] [PubMed] [Google Scholar]
  23. Quinn P. J., Dawson R. M. An analysis of the interaction of protein with lipid monolayers at the air-water interface. Biochem J. 1970 Feb;116(4):671–680. doi: 10.1042/bj1160671. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Quinn P. J., Dawson R. M. Interactions of cytochrome c and [14C]. Biochem J. 1969 Oct;115(1):65–75. doi: 10.1042/bj1150065. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. RIESER P., RIESER C. H. ANABOLIC RESPONSES OF DIAPHRAGM MUSCLE TO INSULIN AND TO OTHER PANCREATIC PROTEINS. Proc Soc Exp Biol Med. 1964 Jul;116:669–671. doi: 10.3181/00379727-116-29339. [DOI] [PubMed] [Google Scholar]
  26. Rodbell M. Metabolism of isolated fat cells. II. The similar effects of phospholipase C (Clostridium perfringens alpha toxin) and of insulin on glucose and amino acid metabolism. J Biol Chem. 1966 Jan 10;241(1):130–139. [PubMed] [Google Scholar]
  27. Turkington R. W. Stimulation of RNA synthesis in isolated mammary cells by insulin and prolactin bound to sepharose. Biochem Biophys Res Commun. 1970 Dec 9;41(5):1362–1367. doi: 10.1016/0006-291x(70)90239-1. [DOI] [PubMed] [Google Scholar]
  28. Vaskovsky V. E., Kostetsky E. Y. Modified spray for the detection of phospholipids on thin-layer chromatograms. J Lipid Res. 1968 May;9(3):396–396. [PubMed] [Google Scholar]
  29. Vilallonga F., Altschul R., Fernandez M. S. Lipid-protein interaction at the air-water interface. Biochim Biophys Acta. 1967 Jul 3;135(3):406–415. doi: 10.1016/0005-2736(67)90030-2. [DOI] [PubMed] [Google Scholar]

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