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Proceedings Paper

Nanosecond Motions Of Genetically-Engineered Antibodies: Structural Elements Controlling Segmental Flexibility Defined By Time-Resolved Emission Anisotropy
Author(s): Theodore G Wensel; William P. Schneider; Vernon T Oi; Lubert Stryel
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Paper Abstract

Immunoglobulins are flexible proteins which display large-amplitude modes of motion on a nanosecond time scale. Different classes of antibodies differ markedly in their degree of segmental flexibility; a number of their essential biological functions are correlated with their nanosecond internal dynamics. These motions can be conveniently monitored by time-resolved fluorescence anisotropy measurements. An instrument built around a synch-pumped cavity-dumped dye laser and a fast time-to-digital convertor with histogramming memory has made it possible to obtain high-quality anisotropy in a few minutes on small amounts (ca. 100 pmol) of protein. Genetic engineering techniques have made it possible to construct a large number of immunoglobulins with identical binding sites for the fluorescent probe dansyllysine. These proteins differ in the heavy chain regions which are responsible for their biological effector functions and their segmental flexibility. We have analyzed a series of such constructs derived by genetic recombination between the genes coding for the mouse isotypes IgG1 and IgG2a. The results identify two regions responsible for their different degrees of segmental flexibility: the hinge region connecting the Fab and Fc portions of the antibodies, and a short stretch (residues 131-139) of sequence in the amino terminal half of the CH1 domain containing five amino acid substitutions.

Paper Details

Date Published: 24 June 1988
PDF: 5 pages
Proc. SPIE 0909, Time-Resolved Laser Spectroscopy in Biochemistry, (24 June 1988); doi: 10.1117/12.945374
Show Author Affiliations
Theodore G Wensel, Stanford University (United States)
William P. Schneider, Stanford University (United States)
Vernon T Oi, Stanford University (United States)
Lubert Stryel, Stanford University (United States)

Published in SPIE Proceedings Vol. 0909:
Time-Resolved Laser Spectroscopy in Biochemistry
Joseph R. Lakowicz, Editor(s)

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