Synthesis and evaluation of fluorescent probes for the detection of calpain activity.
Two new probes for the detection of calpain I activity based on fluorescence resonance energy transfer technology have been synthesized and evaluated. The probes incorporated the cleavage site present in alpha-spectrin, a naturally occurring substrate of calpain I. The design of the internally quenched substrates is such that the calpain-sensitive bond of the peptides (between the Tyr-Gly residues) is located centrally between the donor and the quencher chromophores. The calpain assay protocol is capable of detecting enzymatic activity in the nanomolar region.
Mittoo
Stifun
S
Department of Chemistry, University of Southampton, Highfield, UK.
Sundstrom
Lars E
LE
Bradley
Mark
M
D000595:Amino Acid Sequence; D002154:Calpain / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D004912:Erythrocytes / Q000201:enzymology; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes / Q000138:chemical synthesis* / Q000737:chemistry; D007700:Kinetics; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism; D013049:Spectrin / Q000737:chemistry / Q000235:genetics; D013050:Spectrometry, Fluorescence
D005456:Fluorescent Dyes; D009842:Oligopeptides; D013049:Spectrin; D002154:Calpain