A Plasmodium falciparum dipeptidyl aminopeptidase I participates in vacuolar hemoglobin degradation

被引:146
作者
Klemba, M
Gluzman, I
Goldberg, DE
机构
[1] Washington Univ, Sch Med, Howard Hughes Med Inst, Dept Mol Microbiol, St Louis, MO 63110 USA
[2] Washington Univ, Sch Med, Howard Hughes Med Inst, Dept Med, St Louis, MO 63110 USA
关键词
D O I
10.1074/jbc.M408123200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Intraerythrocytic growth of the human malaria parasite Plasmodium falciparum requires the catabolism of large amounts of host cell hemoglobin. Endoproteolytic digestion of hemoglobin to short oligopeptides occurs in an acidic organelle called the food vacuole. How amino acids are generated from these peptides is not well understood. To gain insight into this process, we have studied a plasmodial ortholog of the lysosomal exopeptidase cathepsin C. The plasmodial enzyme dipeptidyl aminopeptidase 1 (DPAP1) was enriched from parasite extract by two different approaches and was shown to possess hydrolytic activity against fluorogenic dipeptide substrates. To localize DPAP1 we created a transgenic parasite line expressing a chromosomally encoded DPAP1-green fluorescent protein fusion. Green fluorescent protein fluorescence was observed in the food vacuole of live transgenic parasites, and anti-DPAP1 antibody labeled the food vacuole in parasite cryosections. Together these data implicate DPAP1 in the generation of dipeptides from hemoglobin-derived oligopeptides. To assess the significance of DPAP1, we attempted to ablate DPAP1 activity from blood stage parasites by truncating the chromosomal DPAP1-coding sequence. The inability to disrupt the coding sequence indicates that DPAP1 is important for asexual proliferation. The proenzyme form of DPAP1 was found to accumulate in the parasitophorous vacuole of mature parasites. This observation suggests a trafficking route for DPAP1 through the parasitophorous vacuole to the food vacuole.
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页码:43000 / 43007
页数:8
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共 62 条
[11]  
CALLAHAN PX, 1972, FED PROC, V31, P1105
[12]   Stoichiometry and heterogeneity of the pro-region chain in tetrameric human cathepsin C [J].
Cigic, B ;
Krizaj, I ;
Kralj, B ;
Turk, V ;
Pain, RH .
BIOCHIMICA ET BIOPHYSICA ACTA-PROTEIN STRUCTURE AND MOLECULAR ENZYMOLOGY, 1998, 1382 (01) :143-150
[13]   Stable transgene expression in Plasmodium falciparum [J].
Crabb, BS ;
Triglia, T ;
Waterkeyn, JG ;
Cowman, AF .
MOLECULAR AND BIOCHEMICAL PARASITOLOGY, 1997, 90 (01) :131-144
[14]   NUTRITIONAL-REQUIREMENTS OF PLASMODIUM-FALCIPARUM IN CULTURE .1. EXOGENOUSLY SUPPLIED DIALYZABLE COMPONENTS NECESSARY FOR CONTINUOUS GROWTH [J].
DIVO, AA ;
GEARY, TG ;
DAVIS, NL ;
JENSEN, JB .
JOURNAL OF PROTOZOOLOGY, 1985, 32 (01) :59-64
[15]   A SIMPLE METHOD FOR ISOLATING VIABLE MATURE PARASITES OF PLASMODIUM-FALCIPARUM FROM CULTURES [J].
DLUZEWSKI, AR ;
LING, IT ;
RANGACHARI, K ;
BATES, PA ;
WILSON, RJM .
TRANSACTIONS OF THE ROYAL SOCIETY OF TROPICAL MEDICINE AND HYGIENE, 1984, 78 (05) :622-624
[16]   Identification and characterization of falcilysin, a metallopeptidase involved in hemoglobin catabolism within the malaria parasite Plasmodium falciparum [J].
Eggleson, KK ;
Duffin, KL ;
Goldberg, DE .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (45) :32411-32417
[17]   Transformation with human dihydrofolate reductase renders malaria parasites insensitive to WR99210 but does not affect the intrinsic activity of proguanil [J].
Fidock, DA ;
Wellems, TE .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1997, 94 (20) :10931-10936
[18]   A Plasmodium falciparum aminopeptidase gene belonging to the M1 family of zinc-metallopeptidases is expressed in erythrocytic stages [J].
Florent, I ;
Derhy, Z ;
Allary, M ;
Monsigny, M ;
Mayer, R ;
Schrével, J .
MOLECULAR AND BIOCHEMICAL PARASITOLOGY, 1998, 97 (1-2) :149-160
[19]   MOLECULAR CHARACTERIZATION AND INHIBITION OF A PLASMODIUM-FALCIPARUM ASPARTIC HEMOGLOBINASE [J].
FRANCIS, SE ;
GLUZMAN, IY ;
OKSMAN, A ;
KNICKERBOCKER, A ;
MUELLER, R ;
BRYANT, ML ;
SHERMAN, DR ;
RUSSELL, DG ;
GOLDBERG, DE .
EMBO JOURNAL, 1994, 13 (02) :306-317
[20]   Genome sequence of the human malaria parasite Plasmodium falciparum [J].
Gardner, MJ ;
Hall, N ;
Fung, E ;
White, O ;
Berriman, M ;
Hyman, RW ;
Carlton, JM ;
Pain, A ;
Nelson, KE ;
Bowman, S ;
Paulsen, IT ;
James, K ;
Eisen, JA ;
Rutherford, K ;
Salzberg, SL ;
Craig, A ;
Kyes, S ;
Chan, MS ;
Nene, V ;
Shallom, SJ ;
Suh, B ;
Peterson, J ;
Angiuoli, S ;
Pertea, M ;
Allen, J ;
Selengut, J ;
Haft, D ;
Mather, MW ;
Vaidya, AB ;
Martin, DMA ;
Fairlamb, AH ;
Fraunholz, MJ ;
Roos, DS ;
Ralph, SA ;
McFadden, GI ;
Cummings, LM ;
Subramanian, GM ;
Mungall, C ;
Venter, JC ;
Carucci, DJ ;
Hoffman, SL ;
Newbold, C ;
Davis, RW ;
Fraser, CM ;
Barrell, B .
NATURE, 2002, 419 (6906) :498-511