Identification of a processed protein related to the human chaperonins (hsp 60) protein in mammalian kidney.
The chaperonin family of proteins, which includes GroEL protein of E. coli, yeast heat shock protein (hsp-60) and the ribulose-1-5-bisphosphate carboxylase (Rubis Co.) subunit binding protein of plant chloroplasts, shows strong sequence homology to the Chinese hamster ovary (CHO) mitochondrial P1 protein. We have identified a 60 kDa protein from bovine kidney which by N-terminal sequencing gives the amino acid sequence AKDVKFGADARALLMLQGVDLLADA. Bovine whole kidney membranes were delipidated, solubilized with octyl glucoside and fractionated over an affinity column using the amiloride analog 5-N pyrazine amiloride as the ligand. After extensive washing with 200 mM NaCl, the column was eluted with pH 4.0 buffer. Analysis of column fractions on a 7.5% polyacrylamide gel revealed 3-4 bands with a predominant band at 60,000 Da. Amino acid analysis after transfer to immobilon membranes demonstrated sequence identity to the human HSP (60), extending 24 amino acids from the N-terminus, but lacking the leader sequence. These data indicate that a processed form of a protein related to the human HSP (60) chaperonin is associated with a membrane fraction in the mammalian kidney, and that the processed form of the protein binds strongly to an amiloride affinity support.
Ross
W R
WR
Washington University School of Medicine, Department of Medicine, St. Louis, Missouri 63110.
Bertrand
W S
WS
Morrison
A R
AR
D000584:Amiloride / Q000031:analogs & derivatives / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D018833:Chaperonins; D002846:Chromatography, Affinity; D006360:Heat-Shock Proteins / Q000737:chemistry*; D007668:Kidney / Q000737:chemistry*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D011506:Proteins / Q000737:chemistry*; D011719:Pyrazines / Q000378:metabolism
D006360:Heat-Shock Proteins; D010446:Peptide Fragments; D011506:Proteins; D011719:Pyrazines; D000584:Amiloride; D018833:Chaperonins