Isolation and characterization of rat liver mitochondrial ribosomes.
A procedure has been developed that allows characterization of mitochondrial ribosomes and quantitative analysis of the relative composition of constitutive ribosomal proteins. Purified mitochondrial ribosomes were isolated from two rat livers and shown to be active in catalyzing the polymerization of phenylalanine. They differ in sedimentation and spectral properties from cytoplasmic ribosomes isolated from the same livers. The number and relative composition of proteins present in active rat liver mitochondrial ribosomes were investigated using two-dimensional nonequilibrium pH gradient electrophoresis. There were 86 proteins found associated with mitochondrial ribosomes in contrast to 70 proteins found associated with cytoplasmic ribosomes. Comparison of electrophoretic patterns revealed that cytoplasmic ribosomal proteins were considerably more basic than their mitochondrial counterparts. Densitometry demonstrated that the relative changes in the concentrations of these proteins can be measured quantitatively. These procedures, developed for use with two rat livers, allow the rat to be used as an efficient model for further studies into disease states of mitochondrial translation.
Cahill
A
A
Department of Biochemistry, Wake Forest University Medical Center, Winston Salem, North Carolina 27157, USA.
Baio
D L
DL
Cunningham
C C
CC
D000818:Animals; D002458:Cell Fractionation; D002499:Centrifugation, Density Gradient; D003593:Cytoplasm / Q000737:chemistry; D015180:Electrophoresis, Gel, Two-Dimensional; D008297:Male; D008862:Microsomes, Liver / Q000737:chemistry; D008930:Mitochondria, Liver / Q000648:ultrastructure*; D008970:Molecular Weight; D010649:Phenylalanine / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D012269:Ribosomal Proteins / Q000737:chemistry / Q000302:isolation & purification*; D012270:Ribosomes / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D013056:Spectrophotometry, Ultraviolet; D013194:Staining and Labeling
D012269:Ribosomal Proteins; D010649:Phenylalanine