Molecular and functional identification and purification of complement component factor D from urine of patients with chronic renal failure
1990; Elsevier BV; Volume: 27; Issue: 7 Linguagem: Inglês
10.1016/0161-5890(90)90006-l
ISSN1872-9142
AutoresToshio Miyata, Osamu Oda, Reiko Inagi, Satoshi Sugiyama, A. Mlyama, Kenji Maeda, Ichiro Nakashima, Naoki Yamanaka,
Tópico(s)Iron Metabolism and Disorders
ResumoUrine proteins of normal subject and patients with impaired renal function were analyzed by two-dimensional polyacrylamide gel electrophoresis. As a result, a clear spot was detected specifically in urine from patients with obvious renal dysfunction. The isoelectrical point of this unique spot was pH 7.1-7.2 and the flow-rate (Rf) was 0.50-0.55 as that of albumin was 1.0. Partial amino acid sequence analysis revealed that the NH2-terminal to 22nd amino acid sequence was identical with that of complement factor D. We purified 22 mg of this protein (factor D) from 5000 ml of urine from a patient on hemodialysis by three chromatographic steps using DEAE-Sephadex A-50 and Sephacryl S-200. The purified urine factor D gave a single band in sodium dodecyl sulfate polyacrylamide gel electrophoresis at the position of 23 kD, and displayed normal factor D hemolytic activity. The concentrations of factor D estimated by hemolytic assay were 1.9 micrograms/ml of normal serum, less than 0.1 microgram/ml of normal urine, 15 micrograms/ml of patient serum and 50 micrograms/ml of patient urine.
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