Department of Civil and Environmental Engineering, The Center of Advanced Materials for the Purification of Water with Systems, and Department of Pathobiology, University of Illinois at Urbana-Champaign, 205 North Mathews, 3230 Newmark Lab, Urbana, Illinois 61801, USA.
Biomacromolecules. 2010 Aug 9;11(8):2109-15. doi: 10.1021/bm100477j.
We characterized the composition and conformation of Cryptosporidium parvum ( C. parvum ) oocyst wall surface macromolecules and studied their effect on interactions between C. parvum oocyst and quartz surface. Proteinase K and mixed glycosidases were used to modify C. parvum oocyst surface macromolecules. The peptides released by proteinase K and carbohydrates hydrolyzed by mixed glycosidases were respectively analyzed with liquid chromatography/nanoelectrospray ionization tandem mass spectrometry (LC-MS/MS) and phenol-sulfuric acid assay to determine the composition of C. parvum oocyst wall surface macromolecules. Surface potential and polarity of the untreated and proteinase treated C. parvum oocysts revealed information about the conformation of oocyst wall surface macromolecules. The results illustrated that C. parvum oocyst wall is covered by a fluffy layer of glycoproteins. Adhesion kinetics of untreated and proteinase K treated C. parvum oocysts on quartz surface were studied in a radial stagnation point flow cell over a wide range of ionic strength to investigate the effect of C. parvum oocyst wall surface macromolecules on oocysts-quartz interactions. The adhesion rate coefficient of proteinase K treated C. parvum oocysts significantly decreased compared to that of untreated oocysts. This observation indicated that the fluffy layer on C. parvum oocysts wall leads to weaker van der Waals interaction and stronger steric repulsion.
我们对微小隐孢子虫( C. parvum )卵囊壁表面大分子的组成和构象进行了描述,并研究了它们对微小隐孢子虫卵囊与石英表面相互作用的影响。使用蛋白酶 K 和混合糖苷酶来修饰微小隐孢子虫卵囊表面的大分子。用蛋白酶 K 释放的肽和混合糖苷酶水解的碳水化合物分别用液相色谱/纳升电喷雾串联质谱(LC-MS/MS)和苯酚-硫酸法进行分析,以确定微小隐孢子虫卵囊壁表面大分子的组成。未处理和经蛋白酶处理的微小隐孢子虫卵囊的表面电位和极性揭示了卵囊壁表面大分子构象的信息。结果表明,微小隐孢子虫卵囊壁覆盖着一层蓬松的糖蛋白。在广泛的离子强度范围内,在径向停滞点流池内研究了未处理和经蛋白酶 K 处理的微小隐孢子虫卵囊在石英表面上的附着动力学,以研究微小隐孢子虫卵囊壁表面大分子对卵囊-石英相互作用的影响。与未处理的卵囊相比,经蛋白酶 K 处理的微小隐孢子虫卵囊的附着速率系数显著降低。这一观察结果表明,微小隐孢子虫卵囊壁上的蓬松层导致较弱的范德华相互作用和较强的空间排斥。