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 w ith the sequence from the 4197th to 4216th amino acid of perlecan, whi ch is the core protein of the proteoglycan localizing in the systemic capillary basement membranes. It was also found that the molecular wei ght (25000 daltons) of this new protein was comparable to the calculat ed molecular weight of the molecular region of the perlecan from the 4 197th amino acid to the C-terminal. Lastly, it was shown that the anti bodies against this new protein reacted with the perlecan produced by human fibroblasts. All these findings indicated that the new protein i s a perlecan fragment.