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A new carbene based heterobifunctional reagent Photochemical crosslinking of aldolase
Author(s) -
Mogre R.M.,
Batliwala H.F.,
Anjaneyulu P.S.R.,
Lala A.K.
Publication year - 1987
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(87)80965-1
Subject(s) - aldolase a , carbene , reagent , chemistry , photochemistry , polymer chemistry , organic chemistry , enzyme , catalysis
The synthesis of a new photoactivatable heterobifunctional crosslinking reagent, the N ‐oxysuccinimide ester of 2‐carboxy‐9‐diazofluorene, is described. The ability of the parent chromophore 2‐carbomethoxy‐9‐diazofluorene to insert into cyclohexane and methanol has been established. The reagent has been linked to aldolase and the stoichiometry determined. Photolysis of the probe‐linked aldolase indicated that photolysis was very rapid and that the photolysed product was constituted of crosslinked dimer, trimer and tetramer. Increase in concentration of probe linked to aldolase followed by photolysis gave rise to largely tetramer and higher oligomers of aldolase. The use of this carbene‐based reagent vis a vis arylazide‐based reagent for studying protein crosslinking is discussed.