Cysteine-dependent zinc binding by membrane proteins of Giardia lamblia.
The abundant, highly variable surface proteins (VSPs) which cover the surface of Giardia lamblia trophozoites compose a group of extremely cysteine (C)-rich proteins in which more than half of the cysteines are in the motif CXXC. Because of the constancy of these features among the known VSPs and the prominence of cysteine and particularly CXXC in proteins that bind zinc and other metals, we asked whether G. lamblia VSPs bind zinc in vitro. VSPs are the major protein component of Triton X-114 detergent-phase extracts of G. lamblia trophozoites and can be readily identified by surface iodination of intact cells. The partitioning of 65Zn binding into the Triton X-114 detergent phase and the correspondence between surface iodination and zinc binding patterns of four G. lamblia strains or sublines with different VSPs support the idea that VSPs bind zinc. The requirement for renaturation of blots with a reducing agent indicates that Zn2+ is coordinated by cysteine residues, rather than by other amino acids. Binding did not appear to be specific to zinc since it was inhibited by competition with other divalent metal ions. The abundance of the VSPs and the prevalence of metal binding motifs among all known variants suggest that they may play an important role in trophozoite survival and colonization in the host.
Zhang
Y Y
YY
Department of Medicine, Washington University School of Medicine, St. Louis, Missouri 63110.
Aley
S B
SB
Stanley
S L
SL
Gillin
F D
FD
D000818:Animals; D000953:Antigens, Protozoan / Q000378:metabolism*; D000954:Antigens, Surface / Q000378:metabolism*; D002352:Carrier Proteins / Q000032:analysis; D003545:Cysteine / Q000502:physiology*; D016829:Giardia lamblia / Q000378:metabolism*; D015800:Protozoan Proteins; D015032:Zinc / Q000378:metabolism*
D000953:Antigens, Protozoan; D000954:Antigens, Surface; D002352:Carrier Proteins; D015800:Protozoan Proteins; C017800:zinc-binding protein; C072464:variant surface protein H7, Giardia lamblia; D015032:Zinc; D003545:Cysteine