Dynamic laser light scattering in the study of aggregated proteins and in a DNA fragment containing a bend

Albert S. Benight; Donna M. Budzynski; Mohan Amaratunga; Michael J. Lane

doi:10.1117/12.146214

17 June 1993 Dynamic laser light scattering in the study of aggregated proteins and in a DNA fragment containing a bend

Albert S. Benight, Donna M. Budzynski, Mohan Amaratunga, Michael J. Lane

Proceedings Volume 1922, Laser Study of Macroscopic Biosystems; (1993) https://doi.org/10.1117/12.146214
Event: Laser Spectroscopy of Biomolecules: 4th International Conference on Laser Applications in Life Sciences, 1992, Jyvaskyla, Finland

Abstract

We have employed dynamic laser light scattering to investigate the solution behavior of RecA protein from Escherichia coli. RecA is essential for genetic recombination, in vivo, and is the central catalytic protein component of the analogous strand exchange reaction that occurs in vitro between strands of duplex DNA and a homologous single strand. In solution RecA exists in a variety of aggregated forms with sizes and distributions that strongly depend on magnesium concentration, pH, and temperature. With increases in [MgCl₂], temperature or addition of sub-stoichiometric amounts of short duplex DNAs, larger aggregates of RecA are formed. In contrast, increasing pH from 7.3 to 9.0 results in formation of protein aggregates with overall smaller dimensions. Our measurements have also revealed the initial aggregation state of RecA and MgCl₂ induced aggregation depends on the protein preparation procedure. If extreme care is not taken a significant amount of very large, irreversible aggregates contaminate the preparation. These protein species do not display appreciable amounts of magnesium or temperature induced aggregation.

Citation Download Citation

Albert S. Benight, Donna M. Budzynski, Mohan Amaratunga, and Michael J. Lane "Dynamic laser light scattering in the study of aggregated proteins and in a DNA fragment containing a bend", Proc. SPIE 1922, Laser Study of Macroscopic Biosystems, (17 June 1993); https://doi.org/10.1117/12.146214

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