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Binding Dynamics in Biological Systems. Interaction of MOPC-315 with ^(19)F Labelled Nitrophenyl Haptens

Kooistra, Dale A. and Richards, John H. and Smallcombe, Stephen H. (1980) Binding Dynamics in Biological Systems. Interaction of MOPC-315 with ^(19)F Labelled Nitrophenyl Haptens. Organic Magnetic Resonance, 13 (1). pp. 1-8. ISSN 0030-4921. doi:10.1002/mrc.1270130102.

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We have studied the dynamics of binding of ^(19)F labelled haptens by mouse plasmacytoma antibody MOPC-315, a protein which shows specificity for nitrophenyl haptens. The off-rates for the dissociation of hapten from MOPC-315 proteins (7S, Fab′ and Fv) were determined by the application of several methods including a technique which, in certain cases, allows the direct determination of the rate of exchange of nuclei between magnetically non-equivalent sites without requiring prior knowledge of intrinsic line widths. Used in conjunction with independently determined data on line widths, chemical shifts, and binding affinities, these studies show that the rates for hapten association to, or dissociation from, the intact 7S antibody, the Fab′ fragment, and the Fv fragment of MOPC-315 are essentially the same. They also indicate the presence, probably in the Fv region, of a low affinity (K∼ 10^3M^(−1)) site(s) for hapten binding. The mobility of the hapten combining site decreases as the size of the protein increases. These rate data, which were determined at relatively high protein concentrations (up to 40 mg ml^(−1)), agree with on-rates determined at much lower protein concentrations (≤1 mg ml^(−1)); we therefore conclude that protein aggregation, if it does occur, does not significantly affect binding in these systems.

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Additional Information:© 1980 Heyden & Son Ltd. Received 9 June 1978; accepted 30 November 1978. Supported by grant GM-16424 from the National Institute of General Medical Sciences and grant CA-23028 from the National Cancer Institute.
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National Cancer InstituteCA-23028
Issue or Number:1
Record Number:CaltechAUTHORS:20150506-111329487
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Official Citation:Kooistra, D. A., Richards, J. H. and Smallcombe, S. H. (1980), Binding dynamics in biological systems. Interaction of MOPC-315 with 19F labelled nitrophenyl haptens. Org. Magn. Reson., 13: 1–8. doi: 10.1002/mrc.1270130102
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:57274
Deposited By: Ruth Sustaita
Deposited On:06 May 2015 18:24
Last Modified:10 Nov 2021 21:10

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