Purification of a variant-specific surface protein of Giardia lamblia and characterization of its metal-binding properties.
Giardia lamblia, an intestinal parasite of humans and other vertebrates, undergoes surface antigenic variation by modulating the expression of different variant-specific surface proteins (VSP). VSPs are cysteine-rich surface proteins that bind zinc and other heavy metals in vitro. We developed an immunoaffinity chromatographic method to purify a VSP in order to determine its biochemical properties. The sequences of two different proteolytic fragments agreed with the sequence deduced from the cloned gene, and amino-terminal sequence indicated the removal of a 14-residue signal peptide, consistent with the transport of VSP to the cell surface. The protein is not glycosylated and has an isoelectric point of 5.3. X-ray microanalyses indicated that the major metals in Giardia trophozoites, as well as purified VSP, are zinc and iron. The zinc concentration in Giardia cells was found to be 0.43 mM and the iron concentration 0.80 mM when compared with standard samples (zinc) or calculated from a known physical constants (iron). We propose that metal coordination stabilizes VSPs, rendering them resistant to proteolytic attack in the upper small intestine. Moreover, the ability to bind ions by Giardia may play a role in nutritional deficiency and/or malabsorption in heavily infected persons.
Luján
H D
HD
Laboratory of Parasitic Diseases, NIAID, National Institutes of Health, Bethesda, Maryland 20892-0425, USA.
Mowatt
M R
MR
Wu
J J
JJ
Lu
Y
Y
Lees
A
A
Chance
M R
MR
Nash
T E
TE
D000595:Amino Acid Sequence; D000818:Animals; D000953:Antigens, Protozoan / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D000954:Antigens, Surface / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D016829:Giardia lamblia / Q000737:chemistry*; D007501:Iron / Q000378:metabolism*; D008969:Molecular Sequence Data; D015800:Protozoan Proteins; D011817:Rabbits; D015032:Zinc / Q000378:metabolism*
D000953:Antigens, Protozoan; D000954:Antigens, Surface; D015800:Protozoan Proteins; C072464:variant surface protein H7, Giardia lamblia; D007501:Iron; D015032:Zinc