Giardia cyst wall protein 1 is a lectin that binds to curled fibrils of the GalNAc homopolymer.
The infectious and diagnostic stage of Giardia lamblia (also known as G. intestinalis or G. duodenalis) is the cyst. The Giardia cyst wall contains fibrils of a unique beta-1,3-linked N-acetylgalactosamine (GalNAc) homopolymer and at least three cyst wall proteins (CWPs) composed of Leu-rich repeats (CWP(LRR)) and a C-terminal conserved Cys-rich region (CWP(CRR)). Our goals were to dissect the structure of the cyst wall and determine how it is disrupted during excystation. The intact Giardia cyst wall is thin (approximately 400 nm), easily fractured by sonication, and impermeable to small molecules. Curled fibrils of the GalNAc homopolymer are restricted to a narrow plane and are coated with linear arrays of oval-shaped protein complex. In contrast, cyst walls of Giardia treated with hot alkali to deproteinate fibrils of the GalNAc homopolymer are thick (approximately 1.2 microm), resistant to sonication, and permeable. The deproteinated GalNAc homopolymer, which forms a loose lattice of curled fibrils, is bound by native CWP1 and CWP2, as well as by maltose-binding protein (MBP)-fusions containing the full-length CWP1 or CWP1(LRR). In contrast, neither MBP alone nor MBP fused to CWP1(CRR) bind to the GalNAc homopolymer. Recombinant CWP1 binds to the GalNAc homopolymer within secretory vesicles of Giardia encysting in vitro. Fibrils of the GalNAc homopolymer are exposed during excystation or by treatment of heat-killed cysts with chymotrypsin, while deproteinated fibrils of the GalNAc homopolymer are degraded by extracts of Giardia cysts but not trophozoites. These results show the Leu-rich repeat domain of CWP1 is a lectin that binds to curled fibrils of the GalNAc homopolymer. During excystation, host and Giardia proteases appear to degrade bound CWPs, exposing fibrils of the GalNAc homopolymer that are digested by a stage-specific glycohydrolase.
Chatterjee
Aparajita
A
Department of Molecular and Cell Biology, Boston University Goldman School of Dental Medicine, Boston, Massachusetts, USA.
Carpentieri
Andrea
A
Ratner
Daniel M
DM
Bullitt
Esther
E
Costello
Catherine E
CE
Robbins
Phillips W
PW
Samuelson
John
J
D000116:Acetylgalactosamine / Q000378:metabolism*; D002469:Cell Separation; D002473:Cell Wall / Q000737:chemistry / Q000378:metabolism*; D005434:Flow Cytometry; D016829:Giardia lamblia / Q000737:chemistry* / Q000378:metabolism*; D037102:Lectins / Q000378:metabolism; D046529:Microscopy, Electron, Transmission; D015800:Protozoan Proteins / Q000378:metabolism*
C094333:CWP1 protein, Giardia lamblia; D037102:Lectins; D015800:Protozoan Proteins; D000116:Acetylgalactosamine
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