Characterization of protein components of human urinary crystal surface binding substance.
We previously extracted crystal surface binding substance (CSBS) from human urine and showed that it appeared to constitute a substantial proportion of urinary macromolecular inhibitors of calcium oxalate crystallization. CSBS was isolated from human urine and fractionated by three consecutive chromatography procedures in order to characterize protein inhibitors of calcium oxalate crystallization. Sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and NH2-terminal amino acid sequencing revealed that inhibitory fractions eluted from a final, hydroxyapatite column contained prothrombin and osteopontin. Hydroxyapatite column fractions also contained other, unidentified protein inhibitors of calcium oxalate crystallization. CSBS contained also human serum albumin, alpha 1-acid glycoprotein, alpha 1-microglobulin, alpha 2-HS glycoprotein, retinol-binding protein, transferrin, and Tamm-Horsfall protein, but these proteins seemed to play no direct role in inhibitory activity.
Honda
M
M
Department of Urology, Osaka University School of Medicine, Japan.
Yoshioka
T
T
Yamaguchi
S
S
Yoshimura
K
K
Miyake
O
O
Utsunomiya
M
M
Koide
T
T
Okuyama
A
A
D000328:Adult; D000595:Amino Acid Sequence; D002129:Calcium Oxalate / Q000037:antagonists & inhibitors / Q000378:metabolism; D002845:Chromatography; D003460:Crystallization; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000737:chemistry / Q000502:physiology / Q000652:urine*; D006801:Humans; D008297:Male; D053495:Osteopontin; D011516:Prothrombin / Q000032:analysis; D012795:Sialoglycoproteins / Q000032:analysis
D006023:Glycoproteins; C506895:SPP1 protein, human; D012795:Sialoglycoproteins; C029490:nephrocalcin; D053495:Osteopontin; D002129:Calcium Oxalate; D011516:Prothrombin