Purification and partial characterization of an unusual protein of Plasmodium falciparum: histidine-rich protein II.
The human malarial parasite Plasmodium falciparum secretes a histidine-rich protein (HRP-II) from infected erythrocytes. HRP-II has a very high content of histidine (H) (34%), alanine (A) (37%) and aspartic acid (D) (10%) and many contiguous repeats of the sequences AHH and AHHAAD. The histidine content of the protein suggested the potential to bind metal ions. We have demonstrated by metal chelate chromatography an extraordinary capacity of HRP-II to bind zinc ions (Zn2+) and employed this characteristic to isolate the extracellular protein. The HRP-II was further purified by antibody affinity chromatography. The identity of the purified protein was verified by relative molecular weight on denaturing polyacrylamide gels, by reactivity with monoclonal antibodies and monospecific rabbit antiserum, and by comparison of the amino-acid analysis with that derived from the cloned gene sequence. Analysis of the sequence for periodicities using the hydrophobic moment method indicated that HRP-II may potentially form a 3/10 helix. Immunoprecipitation of HRP-II from culture supernatants of parasites metabolically labeled with tritiated sugars showed that the extracellular form of HRP-II is a glycoprotein containing galactose.
Panton
L J
LJ
Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, Bethesda, MD.
McPhie
P
P
Maloy
W L
WL
Wellems
T E
TE
Taylor
D W
DW
Howard
R J
RJ
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000911:Antibodies, Monoclonal; D002241:Carbohydrates / Q000032:analysis; D002614:Chelating Agents; D002846:Chromatography, Affinity; D002942:Circular Dichroism; D003627:Data Interpretation, Statistical; D004591:Electrophoresis, Polyacrylamide Gel; D006031:Glycosylation; D008970:Molecular Weight; D010963:Plasmodium falciparum / Q000032:analysis*; D011487:Protein Conformation; D011506:Proteins / Q000302:isolation & purification* / Q000378:metabolism; D013056:Spectrophotometry, Ultraviolet; D015032:Zinc; D000099053:Histidine-Rich Glycoprotein
D000596:Amino Acids; D000911:Antibodies, Monoclonal; D002241:Carbohydrates; D002614:Chelating Agents; D011506:Proteins; D000099053:Histidine-Rich Glycoprotein; D015032:Zinc