Pattanaik, Priyaranjan and Jain, Bimba and Ravindra, Gudihal and Gopi, Hosahudya N and Pal, Prajna P and Balaram, Hemalatha and Balaram, Padmanabhan (2003) Stage-specific profiling of Plasmodium falciparum proteases using an internally quenched multispecificity protease substrate. In: Biochemical and Biophysical Research Communications, 309 (4). pp. 974-979.
Novel internally quenched fluorescence peptide substrates containing sequence specific sites for cleavage by multiple proteases were designed and synthesized. The 28 and 29 residue peptides contain an N-terminal fluorescence acceptor group, 4-(4-dimethylaminophenylazo) benzoic acid (DABCYL), and a C-terminal fluorescence donor group, 5-(2-aminoethylamino)naphthalene-1-sulfonic acid (EDANS). Efficient energy transfer between the donor and acceptor groups .anking the peptide sequence was achieved by incorporation of a central DPro-Gly segment, which serves as a conformation nucleating site, inducing hairpin formation. This multispecificity protease substrate was used to profile the proteolytic activities in the malarial parasite Plasmodium falciparum in a stage dependent manner using a combination of fluorescence and MALDI mass spectrometry. Cysteine protease activity was shown to be dominating at neutral pH, whereas aspartic protease activity contributed predominantly to the proteolytic repertoire at acidic pH. Maximum proteolysis was observed at the trophozoite stage followed by the schizonts and the rings.
|Item Type:||Journal Article|
|Additional Information:||Copyright for this article belongs to Elsevier Science Ltd|
|Keywords:||Fluorescent protease substrate;Fluorescence resonance energy transfer;beta-Hairpin peptide; Plasmodium falciparum;Malarial proteases;Mass spectrometry|
|Department/Centre:||Division of Biological Sciences > Molecular Biophysics Unit|
|Date Deposited:||30 Jul 2004|
|Last Modified:||19 Sep 2010 04:14|
Actions (login required)