Bovine mitochondrial initiation and elongation factors.
The procedures summarized above provide nearly homogeneous preparations of IF-2mt, EF-Tu. Tsmt, and EF-Gmt. The scheme developed for IF-2mr leads to a 24,000-fold purification of this factor with a 26% recovery of activity. Analysis by SDS-polyacrylamide gel electrophoresis and gel filtration chromatography indicates that this factor functions as a monomer with a molecular weight of about 85,000. The scheme developed EF-Tu.Tsmt provides a 10,000-fold purification with an overall yield of about 10%. The EF-Tumt component in this complex has a molecular weight of about 46,000, whereas EF-Tsmt has a molecular weight of about 32,000 on SDS-polyacrylamide gel electrophoresis. The EF-Tu. Tsmt complex is tightly associated and appears to have a native molecular weight of about 70,000. The five-step purification procedure outlined above for EF-Gmt results in a 14,000-fold purification of EF-Gmt with a 2-5% recovery of activity. Analysis by SDS-polyacrylamide gel electrophoresis and gel filtration chromatography indicates that EF-Gmt functions as a monomeric protein with an apparent molecular weight of about 80,000.
Schwartzbach
C J
CJ
Burroughs Wellcome Company. Research Triangle Park, North Carolina 27709, USA.
Farwell
M
M
Liao
H X
HX
Spremulli
L L
LL
D000818:Animals; D002417:Cattle; D002458:Cell Fractionation / Q000379:methods; D002845:Chromatography / Q000379:methods; D002848:Chromatography, DEAE-Cellulose / Q000379:methods; D002850:Chromatography, Gel / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D004072:Digitonin; D017886:Durapatite; D007202:Indicators and Reagents; D008099:Liver / Q000378:metabolism* / Q000648:ultrastructure; D008930:Mitochondria, Liver / Q000378:metabolism* / Q000648:ultrastructure; D010441:Peptide Chain Elongation, Translational; D010442:Peptide Chain Initiation, Translational; D020652:Peptide Elongation Factor 2; D020653:Peptide Elongation Factor G; D010444:Peptide Elongation Factor Tu / Q000302:isolation & purification* / Q000378:metabolism; D010445:Peptide Elongation Factors / Q000302:isolation & purification* / Q000378:metabolism; D014176:Protein Biosynthesis; D012346:RNA, Transfer, Amino Acyl / Q000378:metabolism; D012270:Ribosomes / Q000378:metabolism* / Q000648:ultrastructure
D007202:Indicators and Reagents; D020652:Peptide Elongation Factor 2; D020653:Peptide Elongation Factor G; D010445:Peptide Elongation Factors; D012346:RNA, Transfer, Amino Acyl; C023512:elongation factor Ts; D017886:Durapatite; D010444:Peptide Elongation Factor Tu; D004072:Digitonin