Simple and high-yield purification of urine protein 1 using immunoaffinity chromatography: evidence for the identity of urine protein 1 and human Clara cell 10-kilodalton protein.
A simple and high-yield purification procedure for urine protein 1 (UP1) using anti-UP1 immunoaffinity chromatography is described. Pure UP1 was obtained in a final yield of 60.2%, and observed as a single electrophoretic band and as a single peak on reversed-phase high-performance liquid chromatography. The N-terminal amino acid residue of UP1 was found to be glutamic acid, contrary to what was reported previously. Furthermore, the N-terminal sequence of UP1 up to 53 amino acids was confirmed to be identical with that of mature human Clara cell 10-kilodalton protein, which inhibits phospholipase A2 activity.
Okutani
R
R
Department of Clinical Pathology, Jichi Medical School, Tochigi-ken, Japan.
Itoh
Y
Y
Hirata
H
H
Kasahara
T
T
Mukaida
N
N
Kawai
T
T
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D007676:Kidney Failure, Chronic / Q000652:urine; D008168:Lung / Q000166:cytology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D011506:Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D011817:Rabbits; D017385:Sequence Homology; D014598:Uteroglobin
D000911:Antibodies, Monoclonal; D011506:Proteins; C556389:SCGB1A1 protein, human; C556390:Scgb1a1 protein, mouse; D014598:Uteroglobin