Small-Angle Neutron Scattering Study of Protein Crowding in Liquid and Solid Phases: Lysozyme in Aqueous Solution, Frozen Solution, and Carbohydrate Powders

Published

July 23, 2012

Author(s)

Joseph E. Curtis, Hirsh Nanda, Sheila Khodadadi, Marcus T. Cicerone, Hyo Jin Lee, Arnold McAuley, Susan Krueger

Abstract

The structure, interactions and inter-protein configurations of the protein lysozyme were studied in a variety of phases that are biotechnologically relevant for proteins using small-angle neutron scattering (SANS) and Monte Carlo simulations. These properties have been studied under a variety of solution conditions before, during, and freezing and freeze-drying in the presence of glucose and trehalose. Contrast variation experiments have also been performed to determine which features of the scattering in the frozen solutions are from the protein and which are from the ice structure. Data from lysozyme at concentrations ranging from 1 mg/mL to 100 mg/mL in solution and water ice with NaCl concentrations ranging from 0 M to 0.4 M are fir to model SANS intensity functions consisting of an ellipsoidal form factor and either a screened ¿Coulomb or hard sphere structure factor. Parameters such as protein volume fraction and long dimension are followed as a function of temperature and salt concentration. The SANS results are compared to real space models of concentrated lysozyme solutions at the same volume fractions obtained from Montel Carlo simulations. A cartoon representation of the frozen lysozyme solution in 0 M NaCl is presented based on the SANS and Monte Carlo results, along with those obtained from other complementary methods.

Citation

Journal of Physical Chemistry B

Volume

116

Pub Type

Journals

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Keywords

small-angle neutron scattering, SANS, protein, interactions, solid phase

Citation

Curtis, J. , , H. , Khodadadi, S. , Cicerone, M. , , H. , McAuley, A. and Krueger, S. (2012), Small-Angle Neutron Scattering Study of Protein Crowding in Liquid and Solid Phases: Lysozyme in Aqueous Solution, Frozen Solution, and Carbohydrate Powders, Journal of Physical Chemistry B, [online], https://tsapps.nist.gov/publication/get_pdf.cfm?pub_id=911025 (Accessed December 8, 2024)