Relative roles of synthesis and degradation in regulating metallothionein accretion
Decay kinetics of {sup 35}S-cysteine (cys) in metallothionein (MT) were used to simultaneously measure rates of MT synthesis and degradation in a HD11 chicken-macrophage cell-line. A reverse-phase (RP) high-performance liquid-chromatography procedure was used to purify 2 MT-isoforms from cytosols with approximately 94% purity. The medium that the macrophages were incubated in was validated to ensure that it contained enough unlabeled cys to adequately chase {sup 35}S-cys released by the degradation of labeled protein. The addition of Zn{sup 2+} and unlabeled cys to the medium did not change the fractional-rates of MT synthesis (FRS) and degradation (FRD). These measurements were also validated by showing that the measured fractional-rate of MT accretion closely approximated the difference between FRS and FRD. When macrophages were incubated in medium supplemented with 50 or 25 {mu}M Zn{sup 2+} the absolute-rate of MT synthesis (ARS) and FRD increased and decreases, respectively. When macrophages were incubated in medium supplemented with 20 or 10 {mu}M Cd{sup 2+}, the ARS increased but the FDR was not changed.
- Research Organization:
- California Univ., Davis, CA (USA)
- OSTI ID:
- 6787562
- Resource Relation:
- Other Information: Thesis (Ph. D.)
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
59 BASIC BIOLOGICAL SCIENCES
CADMIUM
METABOLISM
METALLOTHIONEIN
BIOCHEMISTRY
ZINC
BIOLOGICAL EFFECTS
BIOSYNTHESIS
CHICKENS
FRACTIONATION
LIQUID COLUMN CHROMATOGRAPHY
MACROPHAGES
SULFUR 35
TOXICITY
TRACER TECHNIQUES
ANIMAL CELLS
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIRDS
CHEMISTRY
CHROMATOGRAPHY
CONNECTIVE TISSUE CELLS
DAYS LIVING RADIOISOTOPES
ELEMENTS
EVEN-ODD NUCLEI
FOWL
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
METALLOPROTEINS
METALS
NUCLEI
ORGANIC COMPOUNDS
PHAGOCYTES
PROTEINS
RADIOISOTOPES
SEPARATION PROCESSES
SOMATIC CELLS
SULFUR ISOTOPES
SYNTHESIS
VERTEBRATES
560300* - Chemicals Metabolism & Toxicology
550201 - Biochemistry- Tracer Techniques