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.