Specific adsorption of osteopontin and synthetic polypeptides to calcium oxalate monohydrate crystals.
Protein-crystal interactions are known to be important in biomineralization. To study the physicochemical basis of such interactions, we have developed a technique that combines confocal microscopy of crystals with fluorescence imaging of proteins. In this study, osteopontin (OPN), a protein abundant in urine, was labeled with the fluorescent dye AlexaFluor-488 and added to crystals of calcium oxalate monohydrate (COM), the major constituent of kidney stones. In five to seven optical sections along the z axis, scanning confocal microscopy was used to visualize COM crystals and fluorescence imaging to map OPN adsorbed to the crystals. To quantify the relative adsorption to different crystal faces, fluorescence intensity was measured around the perimeter of the crystal in several sections. Using this method, it was shown that OPN adsorbs with high specificity to the edges between {100} and {121} faces of COM and much less so to {100}, {121}, or {010} faces. By contrast, poly-L-aspartic acid adsorbs preferentially to {121} faces, whereas poly-L-glutamic acid adsorbs to all faces approximately equally. Growth of COM in the presence of rat bone OPN results in dumbbell-shaped crystals. We hypothesize that the edge-specific adsorption of OPN may be responsible for the dumbbell morphology of COM crystals found in human urine.
Taller
Adam
A
Canadian Institutes of Health Research Group in Skeletal Development and Remodeling, and Department of Anatomy and Cell Biology, Schulich School of Medicine and Dentistry, University of Western Ontario, London, Ontario, Canada.
Grohe
Bernd
B
Rogers
Kem A
KA
Goldberg
Harvey A
HA
Hunter
Graeme K
GK
D000327:Adsorption; D002129:Calcium Oxalate / Q000737:chemistry*; D003460:Crystallization; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007669:Kidney Calculi / Q000378:metabolism; D007700:Kinetics; D018613:Microscopy, Confocal; D008855:Microscopy, Electron, Scanning / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods; D053495:Osteopontin / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry*; D011099:Polyglutamic Acid / Q000737:chemistry; D011506:Proteins / Q000737:chemistry; D014556:Urine
D010455:Peptides; D011506:Proteins; D053495:Osteopontin; D011099:Polyglutamic Acid; D002129:Calcium Oxalate