title: Bestatin-based chemical biology strategy reveals distinct roles for malaria M1- and M17-family aminopeptidases.
abstract: Malaria causes worldwide morbidity and mortality, and while chemotherapy remains an excellent means of malaria control, drug-resistant parasites necessitate the discovery of new antimalarials. Peptidases are a promising class of drug targets and perform several important roles during the Plasmodium falciparum erythrocytic life cycle. Herein, we report a multidisciplinary effort combining activity-based protein profiling, biochemical, and peptidomic approaches to functionally analyze two genetically essential P. falciparum metallo-aminopeptidases (MAPs), PfA-M1 and Pf-LAP. Through the synthesis of a suite of activity-based probes (ABPs) based on the general MAP inhibitor scaffold, bestatin, we generated specific ABPs for these two enzymes. Specific inhibition of PfA-M1 caused swelling of the parasite digestive vacuole and prevented proteolysis of hemoglobin (Hb)-derived oligopeptides, likely starving the parasite resulting in death. In contrast, inhibition of Pf-LAP was lethal to parasites early in the life cycle, prior to the onset of Hb degradation suggesting that Pf-LAP has an essential role outside of Hb digestion.
authors.affiliation: Department of Pharmacology, University of Pennsylvania, 433 South University Avenue, 304G Lynch Laboratories, Philadelphia, PA 19104-6018, USA.
authors.forename: Michael B
authors.initials: MB
authors.lastname: Harbut
authors.forename: Geetha
authors.initials: G
authors.lastname: Velmourougane
authors.forename: Seema
authors.initials: S
authors.lastname: Dalal
authors.forename: Gilana
authors.initials: G
authors.lastname: Reiss
authors.forename: James C
authors.initials: JC
authors.lastname: Whisstock
authors.forename: Ozlem
authors.initials: O
authors.lastname: Onder
authors.forename: Dustin
authors.initials: D
authors.lastname: Brisson
authors.forename: Sheena
authors.initials: S
authors.lastname: McGowan
authors.forename: Michael
authors.initials: M
authors.lastname: Klemba
authors.forename: Doron C
authors.initials: DC
authors.lastname: Greenbaum
mesh_terms: D000595:Amino Acid Sequence; D000626:Aminopeptidases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology; D006454:Hemoglobins / Q000378:metabolism; D007930:Leucine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology; D007931:Leucyl Aminopeptidase / Q000037:antagonists & inhibitors; D008288:Malaria / Q000469:parasitology*; D008958:Models, Molecular; D015336:Molecular Probe Techniques; D015335:Molecular Probes / Q000378:metabolism*; D008969:Molecular Sequence Data; D005810:Multigene Family; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D010963:Plasmodium falciparum / Q000187:drug effects / Q000201:enzymology; D040081:Protein Array Analysis; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D013379:Substrate Specificity / Q000187:drug effects
chemical_list: D004791:Enzyme Inhibitors; D006454:Hemoglobins; D015335:Molecular Probes; D019151:Peptide Library; D010455:Peptides; D000626:Aminopeptidases; D007931:Leucyl Aminopeptidase; D007930:Leucine; C012211:ubenimex