Purification and characterization of perlecan fragment in urine of end-stage renal failure patients.
We found a new spot on the two-dimensional electrophoresis pattern of the urine protein from hemodialysis patients. In order to identify the protein forming this new spot, the protein was purified by five steps of chromatography. It was shown that the amino acid sequence of this new protein from the N-terminal to the 20th amino acid was identical with the sequence from the 4197th to 4216th amino acid of perlecan, which is the core protein of the proteoglycan localizing in the systemic capillary basement membranes. It was also found that the molecular weight (25,000 daltons) of this new protein was comparable to the calculated molecular weight of the molecular region of the perlecan from the 4197th amino acid to the C-terminal. Lastly, it was shown that the antibodies against this new protein reacted with the perlecan produced by human fibroblasts. All these findings indicated that the new protein is a perlecan fragment.
Oda
O
O
Bio-Dynamics Research Institute, Nagoya Memorial Hospital, Japan.
Shinzato
T
T
Ohbayashi
K
K
Takai
I
I
Kunimatsu
M
M
Maeda
K
K
Yamanaka
N
N
D000595:Amino Acid Sequence; D015153:Blotting, Western; D002852:Chromatography, Ion Exchange; D015180:Electrophoresis, Gel, Two-Dimensional; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate / Q000737:chemistry / Q000302:isolation & purification / Q000652:urine*; D006801:Humans; D007676:Kidney Failure, Chronic / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry* / Q000652:urine*; D011509:Proteoglycans / Q000737:chemistry / Q000302:isolation & purification / Q000652:urine*; D006435:Renal Dialysis; D017421:Sequence Analysis; D014556:Urine / Q000737:chemistry
D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate; D010446:Peptide Fragments; D011509:Proteoglycans; C071927:perlecan