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Isolation of nuclei from mature peritoneal granulocytes
Author(s) -
Lipke Herbert,
Hester Ralph,
Groff William,
Petrali John
Publication year - 1969
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.1040730202
Subject(s) - cytoplasm , dna , rna , histone , cell nucleus , differential centrifugation , microbiology and biotechnology , nucleoprotein , centrifugation , biochemistry , nucleus , biology , protein biosynthesis , cell , chemistry , gene
Nuclei from mature neutrophil granulocytes were prepared from peritonal exudates of goats. Fluorescein mercuric acetate was required to stabilize the nuclei and fix nucleoproteins. Following differential centrifugation and detergent treatment, electron microscopy showed the interlobar region to be free of cytoplasmic tabs. All of the DNA of the cell was recovered in the nucleus and 71% of the RNA. The DNA : RNA was 6 : 1 in the intact cells, and 9 : 1 in the isolated nuclei. Protein:DNA was 11 : 1 and 4 : 1 for cells and nuclei, respectively. Representative fractions of histones and tryptophan‐rich acidic proteins were prepared with (asp + glu) : (lys + arg + hist) values averaging 0.7 and 1.4 respectively. Histones accounted for 30% of the nuclear proteins while the residual proteins contained the bulk of the cystinyl residues. Granulocytes were characterized by high glycine titers, from 8 to 18% of the nuclear proteins, and 70% of the total free amino acids of the cell.