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Metabolic effects of interleukin 3 on 32D cl23 cells analyzed by NMR
Author(s) -
Hasthorpe S.,
Carver J. A.,
Rees D.,
Campbell I. D.
Publication year - 1987
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.1041330220
Subject(s) - chemistry , biochemistry
31 P NMR of living 32D cl23 cells and 1 H NMR of cell extracts were used to study the metabolic effects of interleukin 3 (IL3). When IL3 was removed from 32D cl23 for 9–10 hours 31 P spectra showed a decrease in sugar phosphate, γATP/ADP, αATP/ADP/NAD, and βATP resonances which declined progressively over a time period of up to 16 hours. By comparison, ATP measurements using the luciferin/luciferase method resulted in the decline of ATP levels from 12 hours in the absence of IL3. At this time, viability of the cells was unaffected. For 1 H NMR experiments cells were grown in the presence and absence of IL3 for 4 and 24 hours, after which acid cell extracts were prepared. These spectra revealed a four‐fold decrease in lactate 4 hours post‐IL3 removal. Alanine levels were unchanged but glycine was elevated 1.5‐fold whilst various other amino acids were elevated slightly. After 24 hours without IL3, only 22% of cells were viable which was reflected in a general decline of most resonance intensities. These findings suggest that IL3 exerts its effect primarily on glucose metabolism and has a delayed secondary effect on maintenance of ATP levels in the cell. We have demonstrated the applicability of high resolution 1 H and 31 P NMR to the study of cellular metabolism in hemopoietic cells.