Title:
Effect of divalent cations and solubilizers in apoferritin and gamma D-crystallin solutions: nucleation, crystallization and light scattering studies

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dc.contributor.advisor Rousseau, Ronald W.
dc.contributor.advisor Sambanis, Athanassios
dc.contributor.author Nwanosike, Quinta M. en_US
dc.contributor.committeeMember Teja, Amyn S.
dc.contributor.committeeMember Nenes, Athanasios
dc.contributor.committeeMember Ingeborg Schmidt-Krey
dc.contributor.department Chemical Engineering en_US
dc.date.accessioned 2010-01-29T19:45:51Z
dc.date.available 2010-01-29T19:45:51Z
dc.date.issued 2009-11-10 en_US
dc.description.abstract Crystallization of proteins in the human body can lead to the development of diseases such as sickle cell anemia and cataract. Understanding protein crystallization can give insight into such diseases. Furthermore, protein crystallization is necessary for protein structure resolution. This is important since resolution of protein structure is the first step towards establishing structure/function relations, and possibly towards performing specific structural modifications that may change the function in desirable directions. Another important application of protein crystallization is in downstream processing in the pharmaceutical industry where it is used for separation and as a final purification step. The present study increases knowledge of interactions between protein molecules during crystallization and hence the crystallization process. Crystallization of proteins in the human body can lead to the development of diseases such as sickle cell anemia and cataract. Understanding the processes involved in protein crystallization can help us gain a better understanding of such diseases. Crystallization of human gamma D-crystallin (HGD) and apoferritin, two proteins found in the lens, was studied in relation to cataract formation. Crystallization of both proteins was studied in the presence of divalent cations which are found at elevated concentrations in cataractous lenses. Results indicate that the divalent cations studied enhance crystallization of these proteins. A thermodynamic property, the osmotic second virial coefficient, was measured in protein solutions and its value was correlated with the occurrence of crystallization. It was found that the second virial coefficient successfully predicted crystallization of both proteins. A new method was developed for indirect measurement of the second virial coefficient using dynamic light scattering. This new method is more robust and efficient than the traditional static light scattering method. Finally the ability of solubilizers to prevent crystallization in HGD solutions was studied. A commercial solubilizer, NDSB-201, was found to increase the energy barrier to nucleation. Although this did not prevent crystallization, it resulted in fewer and smaller crystals being obtained. The naturally occurring alpha A-crystallin was a superior solubilizer to NDSB-201, as it suppressed aggregation and prevented crystallization of HGD under conditions for which NDSB-201 did not. The findings in the present study provide insight into the processes by which protein crystallization occurs and hence into diseases associated with protein crystallization. The findings in the present study provide insight into the processes by which protein crystallization occurs. Using the second virial coefficient to assess whether a protein will crystallize out of solution, approaches for retardation and prevention of protein crystallization, and implications for future research, are discussed. en_US
dc.description.degree Ph.D. en_US
dc.identifier.uri http://hdl.handle.net/1853/31736
dc.publisher Georgia Institute of Technology en_US
dc.subject Light scattering en_US
dc.subject Cataract en_US
dc.subject Second virial coefficient en_US
dc.subject Apoferritin en_US
dc.subject Protein crystallization en_US
dc.subject Gamma D-crystallin en_US
dc.subject.lcsh Light Scattering
dc.subject.lcsh Proteins
dc.subject.lcsh Cations
dc.subject.lcsh Crystal growth
dc.title Effect of divalent cations and solubilizers in apoferritin and gamma D-crystallin solutions: nucleation, crystallization and light scattering studies en_US
dc.type Text
dc.type.genre Dissertation
dspace.entity.type Publication
local.contributor.advisor Rousseau, Ronald W.
local.contributor.advisor Sambanis, Athanassios
local.contributor.corporatename School of Chemical and Biomolecular Engineering
local.contributor.corporatename College of Engineering
relation.isAdvisorOfPublication 54a1d094-6cef-41c7-8168-e36b41eaf4f7
relation.isAdvisorOfPublication 90a12241-15c3-4cbf-bc27-724c028b8fc1
relation.isOrgUnitOfPublication 6cfa2dc6-c5bf-4f6b-99a2-57105d8f7a6f
relation.isOrgUnitOfPublication 7c022d60-21d5-497c-b552-95e489a06569
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