title: Crystallization of calcium oxalates is controlled by molecular hydrophilicity and specific polyanion-crystal interactions.
abstract: To gain more insight into protein structure-function relationships that govern ectopic biomineralization processes in kidney stone formation, we have studied the ability of urinary proteins (Tamm-Horsfall protein, osteopontin (OPN), prothrombin fragment 1 (PTF1), bikunin, lysozyme, albumin, fetuin-A), and model compounds (a bikunin fragment, recombinant-, milk-, bone osteopontin, poly-L-aspartic acid (poly asp), poly-L-glutamic acid (poly glu)) in modulating precipitation reactions of kidney stone-related calcium oxalate mono- and dihydrates (COM, COD). Combining scanning confocal microscopy and fluorescence imaging, we determined the crystal faces of COM with which these polypeptides interact; using scanning electron microscopy, we characterized their effects on crystal habits and precipitated volumes. Our findings demonstrate that polypeptide adsorption to COM crystals is dictated first by the polypeptide's affinity for the crystal followed by its preference for a crystal face: basic and relatively hydrophobic macromolecules show no adsorption, while acidic and more hydrophilic polypeptides adsorb either nonspecifically to all faces of COM or preferentially to {100}/{121} edges and {100} faces. However, investigating calcium oxalates grown in the presence of these polypeptides showed that some acidic proteins that adsorb to crystals do not affect crystallization, even if present in excess of physiological concentrations. These proteins (albumin, bikunin, PTF1, recombinant OPN) have estimated total hydrophilicities from 200 to 850 kJ/mol and net negative charges from -9 to -35, perhaps representing a "window" in which proteins adsorb and coat urinary crystals (support of excretion) without affecting crystallization. Strongest effects on crystallization were observed for polypeptides that are either highly hydrophilic (>950 kJ/mol) and highly carboxylated (poly asp, poly glu), or else highly hydrophilic and highly phosphorylated (native OPN isoforms), suggesting that highly hydrophilic proteins strongly affect precipitation processes in the urinary tract. Therefore, the level of hydrophilicity and net charge is a critical factor in the ability of polypeptides to affect crystallization and to regulate biomineralization processes.
authors.affiliation: CIHR Group in Skeletal Development and Remodeling, School of Dentistry, University of Western Ontario, London, Ontario, Canada.
authors.forename: Bernd
authors.initials: B
authors.lastname: Grohe
authors.forename: Adam
authors.initials: A
authors.lastname: Taller
authors.forename: Peter L
authors.initials: PL
authors.lastname: Vincent
authors.forename: Long D
authors.initials: LD
authors.lastname: Tieu
authors.forename: Kem A
authors.initials: KA
authors.lastname: Rogers
authors.forename: Alexander
authors.initials: A
authors.lastname: Heiss
authors.forename: Esben S
authors.initials: ES
authors.lastname: Sørensen
authors.forename: Silvia
authors.initials: S
authors.lastname: Mittler
authors.forename: Harvey A
authors.initials: HA
authors.lastname: Goldberg
authors.forename: Graeme K
authors.initials: GK
authors.lastname: Hunter
mesh_terms: D000327:Adsorption; D000818:Animals; D000838:Anions / Q000737:chemistry; D002129:Calcium Oxalate / Q000737:chemistry*; D002417:Cattle; D011232:Chemical Precipitation; D003460:Crystallization; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D008855:Microscopy, Electron, Scanning; D011108:Polymers / Q000737:chemistry* / Q000494:pharmacology; D011506:Proteins / Q000737:chemistry / Q000494:pharmacology; D051381:Rats; D013379:Substrate Specificity
chemical_list: D000838:Anions; D011108:Polymers; D011506:Proteins; D002129:Calcium Oxalate