Citric acid extracts a specific set of proteins from isolated cell nuclei.
The treatment of isolated cell nuclei with citric acid was described as a method for separating inner and outer nuclear membrane. Using cell nuclei from bovine cerebral cortex, we can show that citric acid does not cause a separation of the two nuclear membranes, but extracts a specific set of proteins from the nuclei. The extraction of proteins is not just an effect of damaging the nuclear membrane or destructing the cytoskeleton, but rather a specific effect of citric acid treatment. One of the extracted proteins, chosen as a marker for the putative outer nuclear membrane fraction, has an apparent molecular weight of 145 kDa and is located in the nucleoplasm as shown by immunofluorescence microscopy. By sequencing tryptic peptides it was identified as RNA helicase A, an abundant nuclear protein assumed to participate in the processing of mRNA.
Arbeitsgruppe Neurochemie, Institut fur Biochemie, Freie Universitat Berlin, Germany.
U
U
Rosenberger
M
M
Shakibaei
C
C
Weise
P
P
Franke
K
K
Buchner
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D019343:Citric Acid; D007425:Intracellular Membranes / Q000737:chemistry; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000302:isolation & purification*; D020365:RNA Helicases; D012316:RNA Nucleotidyltransferases / Q000302:isolation & purification*
D009687:Nuclear Proteins; D019343:Citric Acid; D012316:RNA Nucleotidyltransferases; D020365:RNA Helicases