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The Journal of Biophysical and Biochemical Cytology logoLink to The Journal of Biophysical and Biochemical Cytology
. 1960 Jul 1;7(4):613–618. doi: 10.1083/jcb.7.4.613

Measurement of Globular Protein Molecules by Electron Microscopy

Cecil E Hall 1
PMCID: PMC2224881  PMID: 14399016

Abstract

A series of molecular species with approximately spherical shape and with molecular weights between 35,000 and 250,000 were shadowed with platinum while resting on a cleaved mica surface. They were backed, stripped from the surface, and examined by electron microscopy. Materials examined were: pepsin, liver alcohol dehydrogenase, yeast alcohol dehydrogenase, glutamic dehydrogenase, polyhedral virus protein (insect), fibrinogen substructure, alkaline phosphatase, and microsomal particles from Escherichia coli. Measurements were made of widths perpendicular to the shadowing direction and heights were deduced from shadow lengths. For those molecular species with well established molecular weights the average heights correlate very well with the diameter of the theoretical sphere but the average widths are too great by 50 to 80 A due to the lateral growth of the deposited metal. Although the distortion in shape of shadowed particles is relatively large, with standardized conditions for shadowing, it is possible to make allowance for the distortion and to obtain reasonably reliable estimates of the dimensions of spherical organic particles down to a molecular weight of about 35,000.

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Selected References

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

  1. FRIEDEN C. Glutamic dehydrogenase. I. The effect of coenzyme on the sedimentation velocity and kinetic behavior. J Biol Chem. 1959 Apr;234(4):809–814. [PubMed] [Google Scholar]
  2. FRIEDEN C. The dissociation of glutamic dehydrogenase by reduced diphosphopyridine nucleotide (DPNH). Biochim Biophys Acta. 1958 Feb;27(2):431–432. doi: 10.1016/0006-3002(58)90364-0. [DOI] [PubMed] [Google Scholar]
  3. GAREN A., LEVINTHAL C. A fine-structure genetic and chemical study of the enzyme alkaline phosphatase of E. coli. I. Purification and characterization of alkaline phosphatase. Biochim Biophys Acta. 1960 Mar 11;38:470–483. doi: 10.1016/0006-3002(60)91282-8. [DOI] [PubMed] [Google Scholar]
  4. HALL C. E. Method for the observation of macromolecules with the electron microscope illustrated with micrographs of DNA. J Biophys Biochem Cytol. 1956 Sep 25;2(5):625–628. doi: 10.1083/jcb.2.5.625. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. HALL C. E., SLAYTER H. S. The fibrinogen molecule: its size, shape, and mode of polymerization. J Biophys Biochem Cytol. 1959 Jan 25;5(1):11–16. doi: 10.1083/jcb.5.1.11. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. HAYES J. E., Jr, VELICK S. F. Yeast alcohol dehydrogenase: molecular weight, coenzyme binding, and reaction equilibria. J Biol Chem. 1954 Mar;207(1):225–244. [PubMed] [Google Scholar]
  7. OLSON J. A., ANFINSEN C. B. The crystallization and characterization of L-glutamic acid dehydrogenase. J Biol Chem. 1952 May;197(1):67–79. [PubMed] [Google Scholar]

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