| Popis: |
When human peripheral blood granulocytes were metabolically labeled at 37 degrees C with [32P]orthophosphate, inorganic polyphosphates became preferentially radiolabeled. Incorporation of radiolabel into the polymer appeared to be ATP-independent. [32P]Polyphosphate was identified by its (i) characteristic lability to acid hydrolysis, (ii) insolubility in barium acetate (pH 4.5), (iii) conversion to [32P]trimetaphosphate, (iv) hydrolysis to [32P]orthophosphate by an exopolyphosphate (Saccharomyces cerevisiae scPPX1), and (v) conversion to a "phosphate ladder" which co-migrated on a polyacrylamide gel with a synthetic phosphate ladder. Also, indirect evidence suggested that the [32P]polyphosphate was strongly, noncovalently associated with another unknown molecule. Particulate fractions (13,000 x g) from lysates of human granulocytes, skin fibroblasts, HL-60 and SK-N-SH cells, all demonstrated radiolabeling of polyphosphate when incubated at 37 degrees C with [32P]orthophosphate. |
