Synthesis and applications of heterobifunctional photocleavable cross-linking reagents.

Abstract:

:This study designed, synthesized, and characterized a number of new heterobifunctional photocleavable cross-linking reagents that may be used to photomodulate the activity of proteins or to prepare caged fluorescent dyes. Biomolecules or fluorophores caged via a thiol group with the BNBASE reagent can be covalently linked to a second protein, ligand, or derivatized surface through the activated carboxyl group. Members of the new class of photocleavable cross-linking reagent can be used to cage amino groups in the molecule of interest, which can then be covalently linked to a second molecule through the thiol-reactive oxirane group. These crosslinking reagents may be used for the following applications: (1) to cage the activity of a protein by masking its active site with a second macromolecule, e.g., aminodextran; (2) to prepare a protein conjugate exhibiting an enhanced or new activity that is lost on irradiation with near-ultraviolet light, e.g., cross-linked actin dimer; (3) to target the caged compound to a specific site by cross linking to a specific antibody; (4) to attach the caged compound to a thiol or amino derivatized surface; and (5) to render the caged compound fluorescent in order to image or to quantify the yield of the photoactivation reaction.

journal_name

Methods Enzymol

journal_title

Methods in enzymology

authors

Marriott G,Ottl J

doi

10.1016/s0076-6879(98)91012-6

subject

Has Abstract

pub_date

1998-01-01 00:00:00

pages

155-75

eissn

0076-6879

issn

1557-7988

pii

S0076-6879(98)91012-6

journal_volume

291

pub_type

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