Abstract
First-order phase transitions of matter, such as condensation and crystallization, proceed through the formation and subsequent growth of ‘critical nuclei’ of the new phase. The thermodynamics and kinetics of the formation of these critical nuclei depend on their structure, which is often assumed to be a compact, three-dimensional arrangement of the constituent molecules or atoms5,6. Recent molecular dynamics simulations have predicted compact nucleus structures for matter made up of building blocks with a spherical interaction field7,8, whereas strongly anisotropic, dipolar molecules may form nuclei consisting of single chains of molecules9. Here we show, using direct atomic force microscopy observations, that the near-critical-size clusters formed during the crystallization of apoferritin, a quasi-spherical protein, and which are representative of the critical nucleus of this system, consist of planar arrays of one or two monomolecular layers that contain 5–10 rods of up to 7 molecules each. We find that these clusters contain between 20 and 50 molecules each, and that the arrangement of the constituent molecules is identical to that found in apoferritin crystals. We anticipate that similarly unexpected critical nucleus structures may be quite common, particularly with anisotropic molecules, suggesting that advanced nucleation theories should treat the critical nucleus structure as a variable.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 51 print issues and online access
$199.00 per year
only $3.90 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Gibbs, J. W. The Collected Works of J.W. Gibbs (Yale Univ. Press, New Haven, 1961).
Oxtoby, D. W. Nucleation of first order phase transitions. Acc. Chem. Res. 31, 91–97 (1998).
Volmer, M. Kinetik der Phasenbildung (Steinkopff, Dresden, 1939 ).
Hill, T. L. Thermodynamics of Small Systems (Benjamin, New York, 1963).
Milchev, A. Electrochemical phase formation on a foreign substrate - basic theoretical concepts and some experimental results. Contemp. Phys. 32, 321–332 (1991).
Chernov, A. A. Modern Crystallography III: Growth of Crystals (Springer, Berlin, 1984).
ten Wolde, P. R. & Frenkel, D. Enhancement of protein crystal nucleation by critical density fluctuations. Science 277, 1975–1978 ( 1997).
Talanquer, V. & Oxtoby, D. W. Crystal nucleation in the presence of a metastable critical point. J. Chem. Phys. 109, 223–227 (1998).
ten Wolde, P. R., Oxtoby, D. W. & Frenkel, D. Coil-globule transition in gas-liquid nucleation of polar fluids. Phys. Rev. Lett. 81, 3695– 3698 (1998).
Harrison, P. M. & Arosio, P. The ferritins: molecular properties, iron storage function and cellular regulation. Biochim. Biophys. Acta 1275, 161–203 (1996).
Lawson, D. M. et al. Solving the structure of human H ferritin by genetically engineering intermolecular crystal contacts. Nature 349, 541–544 (1991).
Hempstead, P. D. et al. Comparison of the three dimensional structures of recombinant human H and horse L ferritins at high resolution. J. Mol. Biol. 268, 424–448 ( 1997).
Thomas, B. R., Carter, D. & Rosenberger, F. Effects of microheterogeneity on horse spleen apoferritin crystallization. J. Cryst. Growth 187, 499 –510 (1997).
Yau, S.-T., Thomas, B. R. & Vekilov, P. G. Molecular mechanisms of crystallisation and defect formation. Phys. Rev. Lett. 85, 353– 356 (2000).
Kuznetsov, Y. G., Malkin, A. J. & McPherson, A. Atomic force microscpopy studies of phase separations in macromolecular systems. Phys. Rev. B 58, 6097–6103 (1998).
Georgalis, Y., Umbach, P., Raptis, J. & Saenger, W. Lysozyme aggregation studied by light scattering. I. Influence of concentration and nature of electrolyte. Acta Crystallogr. D 53, 691– 702 (1997).
Malkin, A. J., Kuznetsov, Y. G. & McPherson, A. Defect structure of macromolecular crystals. J. Struct. Biol. 117, 124–137 (1996 ); Incorporation of microcrystals by growing protein and virus crystals. Proteins Struct. Funct. Genet. 24, 247– 252 (1996).
Kuznetsov, Y. G., Malkin, A. J. & McPherson, A. AFM studies of the nucleation and growth mechanisms of macromolecular crystals. J. Cryst. Growth 196, 489–502 (1999).
Mutaftschiev, B. in Handbook of Crystal Growth (ed. Hurle, D. T. J.) 189– 247 (Elsevier, Amsterdam, 1993).
Kashchiev, D. On the relation between nucleation work, nucleus size, and nucleation rate. J. Chem. Phys. 76, 5098– 5102 (1982).
Oxtoby, D. W. & Kashchiev, D. A general relation between the nucleation work and the size of the nucleus in multicomponent nucleation. J. Chem. Phys. 100, 7665– 7671 (1994).
Stranski, I. N. & Kaischew, R. Über den Mechanismus des Gleichgewichtes kleiner Kriställchen. I. Z. Phys. Chem. B 26, 100–113 (1934).
Kaischew, R. & Stranski, I. N. Über die Thomson-Gibbs’sche Gleichung bei Kristallen. Z. Phys. Chem. B 35, 427–432 (1937).
Vekilov, P. G., Monaco, L. A., Thomas, B. R., Stojanoff, V. & Rosenberger, F. Repartitioning of NaCl and protein impurities in lysozyme crystallization. Acta Crystallogr. D 52, 785–798 (1996).
Galkin, O. & Vekilov, P. G. Direct determination of the nucleation rate of protein crystals. J. Phys. Chem. 103, 10965–10971 (1999); Are nucleation kinetics of protein crystals similar to those of liquid droplets? J. Am. Chem. Soc. 122, 156–163 (2000); Control of protein crystal nucleation around the metastable liquid-liquid phase boundary. Proc. Natl Acad. Sci. USA 97, 6277–6281 ( 2000).
Malkin, A. J. & McPherson, A. Light scattering investigation of the nucleation processes and kinetics of crystallization in macromolecular systems. Acta Crystallogr. D 50, 385– 395 (1994).
Malkin, A. J., Land, T. A., Kuznetsov, Yu. G., McPherson, A. & DeYoreo, J. J. Investigation of virus crystal growth mechanism by in situ atomic force microscopy. Phys. Rev. Lett. 75, 2778–2781 ( 1995).
Petsev, D. N. & Vekilov, P. G. Evidence for non-DLVO hydration interactions in solutions of the protein apoferritin. Phys. Rev. Lett. 84, 1339–1342 ( 2000).
Petsev, D. N., Thomas, B. R., Yau, S.-T. & Vekilov, P. G. Interactions and aggregation of apoferritin molecules in solution: effects of added electrolytes. Biophys. J. 78, 2060 –2069 (2000).
Atkins, P. Physical Chemistry (Freeman, New York, 1998).
Lin, H., Rosenberger, F., Alexander, J. I. D. & Nadarajah, A. Convective-diffusive transport in protein crystal growth. J. Cryst. Growth 151, 153–162 ( 1995).
Acknowledgements
We thank D. W. Oxtoby, D. N. Petsev, J. I. D. Alexander, O. Galkin, S. Weinkauf, J. M. Garcia-Ruiz and N. Booth for suggestions and encouragement; B. R. Thomas for providing pure apoferritin; H. Lin and D. N. Petsev for numerical simulations; and L. Carver for graphics work. This research was supported by the National Heart, Lung, and Blood Institute, NIH, the Life and Microgravity Sciences and Applications Division of NASA, and the State of Alabama through the Centre for Microgravity and Materials Research at the University of Alabama in Huntsville.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Yau, ST., Vekilov, P. Quasi-planar nucleus structure in apoferritin crystallization. Nature 406, 494–497 (2000). https://doi.org/10.1038/35020035
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/35020035
This article is cited by
-
Nucleation of protein mesocrystals via oriented attachment
Nature Communications (2021)
-
Antagonistic cooperativity between crystal growth modifiers
Nature (2020)
-
A mechanism of ferritin crystallization revealed by cryo-STEM tomography
Nature (2020)
-
Quantitative Relationship Between the Nucleation Undercooling of Liquid Iron and Its Liquid Structure: Investigated by In Situ Synchrotron Radiation
Metallurgical and Materials Transactions A (2020)
-
Crystallization tracked atom by atom
Nature (2019)
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.