antibody fluorescence

Bill Melchior, NCTR/FDA wmelchior at NTBTOX.NCTR.FDA.GOV
Wed Sep 23 08:04:08 EST 1992


>Some amino acids (tryptophan and tyrosine) are fluorescent.
>
>If a small molecule is confined to the binding pocket of an antibody,
>it seems that one could, in some circumstances, detect the presence or
>absence of the small molecule by monitoring changes in the fluorescence
>properties of nearby tryptophans and tyrosines. 
>
>In summary, I seek to determine whether it is possible/feasible to
>detect material bound to antibodies by monitoring the fluorescence of
>the antibodies themselves. Any pointers to references would be
>appreciated. Most of the papers I've seen seek to detect the
fluorescence of the bound material, not of the antibody.

Ed-

It's been many years since I've been exposed to this, but I do remember
that it's possible to detect some changes in the environment of aa residues
by fluorescence.  The studies I remember about the binding of small
molecules to proteins, however, I think depended on having a specially designed
small molecule that would quench the fluorescence of the amino acid.

The only reference I could find in my files is old and does not address
your interests directly, but does have some mention of tryptophan fluorescence,
with references -- perhaps it would be a starting point:

MC Chang et al, J Biomolecular Structure and Dynamics, vol 1, 299-318 (1983).
"Internal Dynamics and Overall Motion of Lysozyme Studied by Fluorescence
Depolarization of the Eosin Lysozyme Complex".

Part II of Cantor and Schimmel's 1980 "Biophysical Chemistry" has some
information on the fluorescence properties of the individual aas that might
help you determine the feasiblity of your idea.

Other places to look for relevant papers might include:
 Back issues of the the journal given above.
 The multivolume set "Biomolecular Stereodynamcs", edited by Sarma

Good luck
________________________________________________________________________________
The opinions stated are mine, not those of NCTR or its sponsoring organizations.

Bill Melchior                                ||     OMNISCIENCE
National Center for Toxicological Research   ||    Knowing what
Jefferson, AR  72079                         ||    thou knowest not
(501) 543-7206                               ||    is in a sense
                                             ||    omniscience.
WMELCHIOR at NTDOC.NCTR.FDA.GOV                 ||       from Grooks, Piet Hein



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