Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1977 Nov;74(11):5001–5005. doi: 10.1073/pnas.74.11.5001

Electron microscopy of negatively stained and freeze-etched high density lipoprotein-3 from human serum.

M Ohtsuki, C Edelstein, M Sogard, A M Scanu
PMCID: PMC432086  PMID: 73186

Abstract

High density lipoproteins of d = 1.12 to 1.21 g/ml from human serum (HDL3) were studied by electron microscopy with both negative staining and freeze-etching techniques. For the negatively stained specimens, a modified conventional transmission electron microscope as well as a scanning transmission electron microscope were used. The freeze-etched specimens were examined by a conventional transmission electron microscope. The diameter of HDL3 was found to be 105 +/- 4 A by freeze-etching and 94 +/- 6 A by negative staining. The surface of the HDL3 particles exhibited about 12 discrete domains, 28 +/- 3 A (freeze-etched) and 28 +/- 4 A (negatively stained) in diameter, of undefined chemical composition. Moreover, the freeze-etched specimens revealed an inner core 40 +/- 2 A in diameter, corresponding to estimated values reported previously. All information is consistent with the HDL3 model proposed by B. W. Shen, F. J. Kézdy, and A. M. Scanu [(1977) Proc. Natl. Acad. Sci. USA 74, 837-841], with additional evidence for well-defined surface substructure. The consistency of the images obtained with the various electron microscopy techniques and the marked change in the appearance of the surface in the HDL3 preparations that were digested by phospholipase A2 (EC 3.1.1.4) support the validity of the interpretation.

Full text

PDF
5001

Images in this article

Selected References

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

  1. Forte T., Norum K. R., Glomset J. A., Nichols A. V. Plasma lipoproteins in familial lecithin: cholesterol acyltransferase deficiency: structure of low and high density lipoproteins as revealed by elctron microscopy. J Clin Invest. 1971 May;50(5):1141–1148. doi: 10.1172/JCI106586. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Fukami A., Adachi K. A new method of preparation of a self-perforated micro plastic grid and its application. J Electron Microsc (Tokyo) 1965;14(2):112–118. [PubMed] [Google Scholar]
  3. 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]
  4. Leonard R., Deamer D. W., Armstrong P. Amphibian yolk platelet ultrastructure visualized by freeze-etching. J Ultrastruct Res. 1972 Jul;40(1):1–24. doi: 10.1016/s0022-5320(72)80019-4. [DOI] [PubMed] [Google Scholar]
  5. Nichols A. V. Functions and interrelationships of different classes of plasma lipoproteins. Proc Natl Acad Sci U S A. 1969 Nov;64(3):1128–1137. doi: 10.1073/pnas.64.3.1128. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Pattnaik N. M., Kézdy F. J., Scanu A. M. Kinetic study of the action of snake venom phospholipase A2 on human serum high density lipoprotein 3. J Biol Chem. 1976 Apr 10;251(7):1984–1990. [PubMed] [Google Scholar]
  7. Scanu A. Forms of human serum high density lipoprotein protein. J Lipid Res. 1966 Mar;7(2):295–306. [PubMed] [Google Scholar]
  8. Shen B. W., Scanu A. M., Kézdy F. J. Structure of human serum lipoproteins inferred from compositional analysis. Proc Natl Acad Sci U S A. 1977 Mar;74(3):837–841. doi: 10.1073/pnas.74.3.837. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Steere R. L. Freeze-etching and direct observation of freezing damage. Cryobiology. 1969 Nov-Dec;6(3):137–150. doi: 10.1016/s0011-2240(69)80344-5. [DOI] [PubMed] [Google Scholar]
  10. Tardieu A., Mateu L., Sardet C., Weiss B., Luzzati V. Structure of human serum lipoproteins in solution. II. Small-angle x-ray scattering study of HDL and LDL. J Mol Biol. 1976 Feb 25;101(2):129–153. doi: 10.1016/0022-2836(76)90368-5. [DOI] [PubMed] [Google Scholar]
  11. Verdery R. B., Nichols A. V. Arrangement of lipid and protein in human serum high density lipoproteins: a proposed model. Chem Phys Lipids. 1975 Apr;14(2):123–134. doi: 10.1016/0009-3084(75)90054-7. [DOI] [PubMed] [Google Scholar]
  12. Wall J., Langmore J., Isaacson M., Crewe A. V. Scanning transmission electron microscopy at high resolution. Proc Natl Acad Sci U S A. 1974 Jan;71(1):1–5. doi: 10.1073/pnas.71.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Wells M. A., Hanahan D. J. Studies on phospholipase A. I. Isolation and characterization of two enzymes from Crotalus adamanteus venom. Biochemistry. 1969 Jan;8(1):414–424. doi: 10.1021/bi00829a057. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES