Search

Your search keyword '"Lackovic K"' showing total 2 results
Start Over Author "Lackovic K" Publisher american chemical society
2 results on '"Lackovic K"'

1. Transition state mimetics of the Plasmodium export element are potent inhibitors of Plasmepsin V from P. falciparum and P. vivax.

Author

Sleebs BE, Gazdik M, O'Neill MT, Rajasekaran P, Lopaticki S, Lackovic K, Lowes K, Smith BJ, Cowman AF, and Boddey JA

Subjects
Aspartic Acid Endopeptidases chemistry, Aspartic Acid Endopeptidases metabolism, Cell Line, Drug Discovery, Erythrocytes drug effects, Erythrocytes parasitology, Humans, Models, Molecular, Protein Conformation, Proteolysis drug effects, Structure-Activity Relationship, Aspartic Acid Endopeptidases antagonists & inhibitors, Biomimetic Materials pharmacology, Plasmodium falciparum drug effects, Plasmodium falciparum enzymology, Plasmodium vivax drug effects, Plasmodium vivax enzymology, Protease Inhibitors pharmacology
Abstract

Following erythrocyte invasion, malaria parasites export a catalogue of remodeling proteins into the infected cell that enable parasite development in the human host. Export is dependent on the activity of the aspartyl protease, plasmepsin V (PMV), which cleaves proteins within the Plasmodium export element (PEXEL; RxL↓xE/Q/D) in the parasite's endoplasmic reticulum. Here, we generated transition state mimetics of the native PEXEL substrate that potently inhibit PMV isolated from Plasmodium falciparum and Plasmodium vivax. Through optimization, we identified that the activity of the mimetics was completely dependent on the presence of P1 Leu and P3 Arg. Treatment of P. falciparum-infected erythrocytes with a set of optimized mimetics impaired PEXEL processing and killed the parasites. The striking effect of the compounds provides a clearer understanding of the accessibility of the PMV active site and reaffirms the enzyme as an attractive target for the design of future antimalarials.

Published
2014
Full Text
View/download PDF

2. A biosensor of SRC family kinase conformation by exposable tetracysteine useful for cell-based screening.

Author

Irtegun S, Wood R, Lackovic K, Schweiggert J, Ramdzan YM, Huang DC, Mulhern TD, and Hatters DM

Subjects
Amino Acid Sequence, Animals, COS Cells, Cell Line, Chlorocebus aethiops, Coloring Agents chemistry, Coloring Agents metabolism, Cysteine metabolism, Drug Evaluation, Preclinical methods, Humans, Models, Molecular, Protein Conformation drug effects, Protein Kinase Inhibitors pharmacology, src Homology Domains drug effects, src-Family Kinases antagonists & inhibitors, src-Family Kinases metabolism, Biosensing Techniques methods, Cysteine chemistry, src-Family Kinases chemistry
Abstract

We developed a new approach to distinguish distinct protein conformations in live cells. The method, exposable tetracysteine (XTC), involved placing an engineered tetracysteine motif into a target protein that has conditional access to biarsenical dye binding by conformational state. XTC was used to distinguish open and closed regulatory conformations of Src family kinases. Substituting just four residues with cysteines in the conserved SH2 domain of three Src-family kinases (c-Src, Lck, Lyn) enabled open and closed conformations to be monitored on the basis of binding differences to biarsenical dyes FlAsH or ReAsH. Fusion of the kinases with a fluorescent protein tracked the kinase presence, and the XTC approach enabled simultaneous assessment of regulatory state. The c-Src XTC biosensor was applied in a boutique screen of kinase inhibitors, which revealed six compounds to induce conformational closure. The XTC approach demonstrates new potential for assays targeting conformational changes in key proteins in disease and biology.

Published
2014
Full Text
View/download PDF

Catalog

Books, media, physical & digital resources
See catalog results