TY - JOUR
T1 - Investigation of human lymphocyte plasma membrane associated nucleic acid
AU - Melera, Peter W.
AU - Cronin-Sheridan, A. P.
N1 - Funding Information:
The authors acknowledge the cooperation of Dr. L.F. Cavalieri in whose laboratory this work was carried out. We also thank Ms. J. Murcott for excellent technical assistance and Dr. R.H.F. Peterson for many helpful discussions. This work was supported in part by NCI grant CA-08748 and AEC contract AT(11-1)-3521).
PY - 1976/5/19
Y1 - 1976/5/19
N2 - Plasma membranes were prepared from the human lymphocyte cell line WIL23A by hypotonic swelling, Dounce homogenization, differential and equilibrium centrifugation. The resulting vesiculated membrane fragments were found to have densities of 1.10 and 1.17 g/ml, and were defined by lactoperoxidase mediated whole cell iodination, l-[3H]fucose incorporation, 5′-nucleotidase activity (EC 3.1.3.5) and electron micrographic visualization. Recovery of plasma membrane from whole cell homogenates was estimated to be approximately 30-35% as judged by the recovery of 125I-labeled cell surface protein. When plasma membranes were prepared from cells which had been incubated for 18 h in the presence of 0.5 μCi/ml [3H] thymidine such that greater than 109 acid insoluble counts could be demonstrated in the whole cell h homogenates, no [3H] thymidine label and presumably, therefore, no DNA, could be shown to be coincident with either the 1.10 or 1.17 density. Similar experiments with [3H] uridine suggested that 90% of the plasma membranes did not contain RNA, while 10% remained questionable.
AB - Plasma membranes were prepared from the human lymphocyte cell line WIL23A by hypotonic swelling, Dounce homogenization, differential and equilibrium centrifugation. The resulting vesiculated membrane fragments were found to have densities of 1.10 and 1.17 g/ml, and were defined by lactoperoxidase mediated whole cell iodination, l-[3H]fucose incorporation, 5′-nucleotidase activity (EC 3.1.3.5) and electron micrographic visualization. Recovery of plasma membrane from whole cell homogenates was estimated to be approximately 30-35% as judged by the recovery of 125I-labeled cell surface protein. When plasma membranes were prepared from cells which had been incubated for 18 h in the presence of 0.5 μCi/ml [3H] thymidine such that greater than 109 acid insoluble counts could be demonstrated in the whole cell h homogenates, no [3H] thymidine label and presumably, therefore, no DNA, could be shown to be coincident with either the 1.10 or 1.17 density. Similar experiments with [3H] uridine suggested that 90% of the plasma membranes did not contain RNA, while 10% remained questionable.
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U2 - 10.1016/0005-2787(76)90139-8
DO - 10.1016/0005-2787(76)90139-8
M3 - Article
C2 - 1268257
AN - SCOPUS:0017153795
SN - 0005-2787
VL - 432
SP - 300
EP - 311
JO - BBA Section Nucleic Acids And Protein Synthesis
JF - BBA Section Nucleic Acids And Protein Synthesis
IS - 3
ER -