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{sup 3}H-tetracycline as a proxy for {sup 41}Ca for measuring dietary perturbations of bone resorption  

Energy Technology Data Exchange (ETDEWEB)

Our group is interested in evaluating early effects of dietary interventions on bone loss. Postmenopausal women lose bone following reduction in estrogen which leads to increased risk of fracture. Traditional means of monitoring bone loss and effectiveness of treatments include changes in bone density, which takes 6 months to years to observe effects, and changes in biochemical markers of bone turnover, which are highly variable and lack specificity. Prelabeling bone with {sup 41}Ca and measuring urinary {sup 41}Ca excretion with accelerator mass spectrometry provides a sensitive, specific, and rapid approach to evaluating effectiveness of treatment. To better understand {sup 41}Ca technology as a tool for measuring effective treatments on reducing bone resorption, we perturbed bone resorption by manipulating dietary calcium in rats. We used {sup 3}H-tetracycline ({sup 3}H-TC) as a proxy for {sup 41}Ca and found that a single dose is feasible to study bone ...

2007-06-15

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Identification of proteins important for tetracycline (TC) binding to ribosomes by single protein omission reconstitution (SPORE) experiments  

Energy Technology Data Exchange (ETDEWEB)

TC inhibits protein synthesis in E. coli by interfering with aminoacyl-tRNA binding to the ribosomal A site, and there is strong evidence that such inhibition results from TC binding to a high affinity site on the 30S subunit. The SPORE approach has been used to define those 30S proteins that are potentially important for high affinity TC binding, measured as the (/sup 3/H)-TC co-sedimenting with the reconstitution particle through a sucrose density gradient. Reverse phase-HPLC has been used both to prepare ribosomal proteins and to analyze the protein content of reconstituted particles. The results obtained so far show that protein S7, as well as some proteins linked to S7 in the 30S assembly map, are important for TC binding, whereas other ribosomal proteins are not. These results are in very good accord with their earlier photoaffinity labeling studies that strongly implicated S7 as forming part of the TC binding site. Interestingly, protein S18, which is ...

1987-05-01