Four plasmepsins are active in the Plasmodium falciparum food vacuole, including a protease with an active-site histidine

Hemoglobin degradation is a metabolic process that is central to the growth and maturation of the malaria parasite Plasmodium falciparum. Two aspartic proteases that initiate degradation, plasmepsins (PMs) I and II, have been identified and extensively characterized. Eight additional PM genes are present in the P. falciparum genome. To better understand the enzymology of hemoglobin degradation, it is necessary to determine which of these genes are expressed when hemoglobin degradation is occurring, which encode active enzymes, and which gene products are found in the food vacuole where catabolism takes place. Our genome-wide analysis reveals that PM I, II, and IV and histo-aspartic protease encode hemoglobin-degrading food vacuole proteases. Despite having a histidine in place of one of the catalytic aspartic acids conserved in other aspartic proteases, histo-aspartic protease is an active hydrolase.

[1]  R. Roberts,et al.  The diversity and evolutionary relationships of the pregnancy-associated glycoproteins, an aspartic proteinase subfamily consisting of many trophoblast-expressed genes. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[2]  R. Roberts,et al.  Identification of the major pregnancy-specific antigens of cattle and sheep as inactive members of the aspartic proteinase family. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[3]  J T Palmer,et al.  Antimalarial effects of vinyl sulfone cysteine proteinase inhibitors , 1996, Antimicrobial agents and chemotherapy.

[4]  P. Rosenthal,et al.  Inhibition of a Plasmodium vinckei cysteine proteinase cures murine malaria. , 1993, The Journal of clinical investigation.

[5]  D. Goldberg,et al.  Aspartic proteases of Plasmodium falciparum and other parasitic protozoa as drug targets. , 2001, Trends in parasitology.

[6]  Dennis E. Kyle,et al.  New Class of Small Nonpeptidyl Compounds BlocksPlasmodium falciparum Development In Vitro by Inhibiting Plasmepsins , 2001, Antimicrobial Agents and Chemotherapy.

[7]  I. Gluzman,et al.  Kinetic analysis of plasmepsins I and II aspartic proteases of the Plasmodium falciparum digestive vacuole. , 1996, Molecular and biochemical parasitology.

[8]  D. Goldberg,et al.  Biosynthesis and Maturation of the Malaria Aspartic Hemoglobinases Plasmepsins I and II* , 1997, The Journal of Biological Chemistry.

[9]  G. Reddy,et al.  Sequence, expression and modeled structure of an aspartic proteinase from the human malaria parasite Plasmodium falciparum. , 1994, Molecular and biochemical parasitology.

[10]  I. Gluzman,et al.  Characterization of native falcipain, an enzyme involved in Plasmodium falciparum hemoglobin degradation. , 1996, Molecular and biochemical parasitology.

[11]  D. Bur,et al.  The aspartic proteinase from the rodent parasite Plasmodium berghei as a potential model for plasmepsins from the human malaria parasite, Plasmodium falciparum , 1999, FEBS letters.

[12]  P. Rosenthal,et al.  A malarial cysteine proteinase is necessary for hemoglobin degradation by Plasmodium falciparum. , 1988, The Journal of clinical investigation.

[13]  I. Gluzman,et al.  Identification of potent inhibitors of Plasmodium falciparum plasmepsin II from an encoded statine combinatorial library. , 1998, Bioorganic & medicinal chemistry letters.

[14]  D. Goldberg,et al.  Identification and Characterization of Falcilysin, a Metallopeptidase Involved in Hemoglobin Catabolism within the Malaria Parasite Plasmodium falciparum* , 1999, The Journal of Biological Chemistry.

[15]  U. Certa,et al.  A distinct member of the aspartic proteinase gene family from the human malaria parasite Plasmodium falciparum , 1999, FEBS letters.

[16]  J. Ravel,et al.  Radiolabeling of proteins by reductive alkylation with [14C]formaldehyde and sodium cyanoborohydride. , 1978, Analytical biochemistry.

[17]  Irwin W. Sherman,et al.  Malaria : parasite biology, pathogenesis, and protection , 1998 .

[18]  D. Chakrabarti,et al.  Antimalarial Activities of Polyhydroxyphenyl and Hydroxamic Acid Derivatives , 1998, Antimicrobial Agents and Chemotherapy.

[19]  J. E. Celis,et al.  Cell Biology: A Laboratory Handbook , 1997 .

[20]  D. Goldberg,et al.  Hemoglobin degradation in Plasmodium-infected red blood cells. , 1993, Seminars in cell biology.

[21]  P. Rosenthal,et al.  Isolation and characterization of a cysteine proteinase gene of Plasmodium falciparum. , 1992, Molecular and biochemical parasitology.

[22]  E. Harlow,et al.  Using Antibodies: A Laboratory Manual , 1999 .

[23]  P. Rosenthal.  Antimalarial chemotherapy: mechanisms of action, resistance, and new directions in drug discovery. , 2002 .

[24]  P. Rosenthal,et al.  Proteases of protozoan parasites. , 1999, Advances in parasitology.

[25]  B. Chait,et al.  Hemoglobin degradation in the human malaria pathogen Plasmodium falciparum: a catabolic pathway initiated by a specific aspartic protease , 1991, The Journal of experimental medicine.

[26]  Robert A. Copeland,et al.  Enzymes: A Practical Introduction to Structure, Mechanism, and Data Analysis , 1996 .

[27]  P. Rosenthal,et al.  Antimalarial effects in mice of orally administered peptidyl cysteine protease inhibitors. , 1999, Bioorganic & medicinal chemistry.

[28]  W. Trager,et al.  Human malaria parasites in continuous culture. , 1976, Science.

[29]  J. Beckers,et al.  The gene encoding bovine pregnancy-associated glycoprotein-1, an inactive member of the aspartic proteinase family. , 1995, Gene.

[30]  U. Certa,et al.  Expression and characterisation of plasmepsin I from Plasmodium falciparum. , 1997, European journal of biochemistry.

[31]  I. Gluzman,et al.  Molecular characterization and inhibition of a Plasmodium falciparum aspartic hemoglobinase. , 1994, The EMBO journal.

[32]  Neil D. Rawlings,et al.  Handbook of proteolytic enzymes , 1998 .

[33]  I. Gluzman,et al.  Order and specificity of the Plasmodium falciparum hemoglobin degradation pathway. , 1994, The Journal of clinical investigation.

[34]  T. Boulikas,et al.  Silver staining of proteins in polyacrylamide gels. , 1981, Analytical biochemistry.

[35]  J. Vennerstrom.  Antimalarial Chemotherapy: Mechanisms of Action, Resistance, and New Directions in Drug Discovery Edited by P. J. Rosenthal. Humana Press, Totowa, NJ. 2001. xi + 396 pp. 18 × 26 cm. ISBN 0-89603-670-7. $135.00. , 2001 .

[36]  T. Blundell,et al.  Comparative modelling and analysis of amino acid substitutions suggests that the family of pregnancy-associated glycoproteins includes both active and inactive aspartic proteinases. , 1996, Protein engineering.

[37]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.