Pritzker School of Molecular Engineering, University of Chicago, Chicago, Illinois, United States of America.
Department of Urology, Department of Veterans Affairs Medical Center and the Medical College of Wisconsin, Milwaukee, Wisconsin, United States of America.
PLoS One. 2021 Sep 23;16(9):e0257515. doi: 10.1371/journal.pone.0257515. eCollection 2021.
Despite the apparent importance of matrix proteins in calcium oxalate kidney stone formation, the complexity of the protein mixture continues to elude explanation. Based on a series of experiments, we have proposed a model where protein aggregates formed from a mixture containing both strongly charged polyanions and strongly charged polycations could initiate calcium oxalate crystal formation and crystal aggregation to create a stone. These protein aggregates also preferentially adsorb many weakly charged proteins from the urine to create a complex protein mixture that mimics the protein distributions observed in patient samples. To verify essential details of this model and identify an explanation for phase selectivity observed in weakly charged proteins, we have examined primary structures of major proteins preferring either the matrix phase or the urine phase for their contents of aspartate, glutamate, lysine and arginine; amino acids that would represent fixed charges at normal urine pH of 6-7. We verified enrichment in stone matrix of proteins with a large number of charged residues exhibiting extreme isoelectric points, both low (pI<5) and high (pI>9). We found that the many proteins with intermediate isoelectric points exhibiting preference for stone matrix contained a smaller number of charge residues, though still more total charges than the intermediate isoelectric point proteins preferring the urine phase. While other sources of charge have yet to be considered, protein preference for stone matrix appears to correlate with high total charge content.
尽管基质蛋白在草酸钙肾结石形成中显然很重要,但蛋白质混合物的复杂性仍难以解释。基于一系列实验,我们提出了一个模型,即在含有强带电荷的聚阴离子和强带电荷的聚阳离子的混合物中形成的蛋白质聚集体可以启动草酸钙晶体的形成和晶体聚集,从而形成结石。这些蛋白质聚集体还优先从尿液中吸附许多带弱电荷的蛋白质,形成一种复杂的蛋白质混合物,模拟在患者样本中观察到的蛋白质分布。为了验证该模型的重要细节,并为弱电荷蛋白质中观察到的相选择性找到解释,我们检查了主要蛋白质的一级结构,这些蛋白质根据其天冬氨酸、谷氨酸、赖氨酸和精氨酸的含量,优先选择基质相或尿液相;在正常尿液 pH 值为 6-7 时,这些氨基酸代表固定电荷。我们验证了在结石基质中富含大量带电荷残基的蛋白质,这些残基具有极端等电点,既低(pI<5)又高(pI>9)。我们发现,许多具有中间等电点、优先与结石基质结合的蛋白质含有较少的电荷残基,但总电荷数仍多于优先与尿液相结合的中间等电点蛋白质。虽然还需要考虑其他电荷来源,但蛋白质对结石基质的偏好似乎与高总电荷含量有关。