Abstract
Structural information on clathrin coated vesicles has been obtained by small angle neutron scattering using contrast variation. A characteristic peak in the neutron scattering profile, which is apparent in 75 % D2O, as well as in H2O, disappears when contrast matching the protein component of the coated vesicles in 42% D2O. Neutron, as well as dynamic, light scattering give a coated vesicle size of about 900 Å in H2O and D2O, but for neutron scattering the diameter decreases when matching out the protein coat of the clathrin coated vesicles. From the match point for the clathrin coated vesicles it is demonstrated that the clathrin cages do contain internal membrane. The mass of 34 MD and composition of 75% protein and 25% lipid found from the analysis of the small-angle scattering data are both in good agreement with the values reported in the literature. Electron microscopy gives an average outer diameter of 880 Å for the coated vesicles and an average diameter of 460 Å for the vesicle itself.
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References
Bauer R, Behan M, Jones G, Hansen S, Mortensen K, Særmark T, Øgendal L (1991) Small angle scattering studies on clathrin-coated vesicles. J Appl Crystallogr 24:815–821
Bevington BR (1969) Data reduction and error analysis for the physical sciences. McGraw-Hill, New York
Cusack S, Ruigrok RWH, Krygsman PCJ, Mellema JE (1985) Structure and composition of influenza virus. A small-angle neutron scattering study. J Mol Biol 186:565–582
Crowther RA, Finch JT, Pearse BMF (1976) On the structure of coated vesicles. J Mol Biol 103:785–798
Dubochet J, Lepault J, Freeman R, Berriman JA, Homo JC (1976) Electron microscopy of frozen water and aqueous solutions. J Microsc 128:219–237
Glatter O (1977) A new method for the evaluation of small-angle scattering data. J Appl Crystallogr 10:415–421
Glatter O (1982) in: Glatter O, Kratky O (eds) Small angle X-ray scattering. Academic Press, London
Hansen S, Skov Pedersen J (1991) A comparison of three different methods for analysing small-angle scattering data. J Appl Crystallogr 24:514–518
Ibel K, Stuhrmann HB (1975) Comparison of neutron and X-ray scattering of dilute myoglobin solutions. J Mol Biol 93:255–265
Jacrot B, Zacchai G (1981) Determination of molecular weight by neutron scattering Biopolymers 20:2413–2426
Moore PB (1980) Small-angle scattering. Information content and error analysis. J Appl Crystallogr 13:168–175
Pearse BMF (1975) Coated vesicles from pig brain: Purification and biochemical characterization. J Mol Biol 97:93–98
Pearse BMF, Crowther RA (1987) Structure and assembly of coated vesicles. Ann Rev Biophys Chem 16:49–68
Skov Pedersen J, Posselt D, Mortensen K (1990) Analytical treatment of the resolution function for small-angle scattering. J Appl Crystallogr 23:321–333
Vigers GPA, Crowther RA, Pearse BMF (1986) Location of the 100 kD-50 kD accessory proteins in clathrin coats. EMBO J 5:2079–2085
Winkler FK, Stanley KK (1983). Clathrin heavy chain, light chain interactions. EMBO J 2:1393–1400
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Bauer, R., Behan, M., Clarke, D. et al. Contrast variation studies of clathrin coated vesicles by small-angle neutron scattering. Eur Biophys J 21, 129–136 (1992). https://doi.org/10.1007/BF00185427
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DOI: https://doi.org/10.1007/BF00185427