It has recently been demonstrated that the chemotactic peptide N-formyl-Met-Leu-Phe activates phospholipase D (PLD) in dimethyl sulfoxide-differentiated HL-60 granulocytes to produce phosphatidic acid (PA) and, in the presence of ethanol, phosphatidylethanol (PEt) (Pai, J.-K., Siegel, M. I., Egan, R. W., and Billah, M. M. (1988) J. Biol. Chem. 263, 12472-12477). We now report that biologically ...

In order to obtain a highly concentrated, pure and super-hot [ƒÁ-32P]ATP, we improved the purification method of super-hot [ƒÁ-32P]ATP which was synthesized by the method of Johnson and Walseth (1979). The super-hot [ƒÁ-32P]ATP was synthesized in a relatively large volume (2 ml) of reaction mixture and purified using semi-micro scale anion exchange chromatography (Dowex 1•~2, 60-70ƒÊl column vo...

Export of mRNA from the nucleus to the cytoplasm was studied in mature Xenopus laevis oocytes. In vitro transcribed, capped 32P-labeled mRNA was microinjected into nuclei, and its appearance in the cytoplasm measured by counting radioactivity or by RNA extraction and gel electrophoresis. Both for a 5.0-kb transferrin receptor mRNA and a 2.0-kb 4F2 antigen heavy chain mRNA we found saturable tra...

Upon illumination of spinach chloroplasts in the presence of [32P]orthophosphate and AMP and absence of substrate ADP, coupling factor-bound [32P]ADP is formed. This bound r3’P]ADP is stable enough to withstand purification of the coupling factor to homogeneity. However, the ability of this specially bound [““PIADP to serve as the ultimate phosphoryl donor to substrate ADP for the synthesis of ...

Equilibration of the phosphorus in radioactive phosphoric acid with the phosphorus in phosphorus oxychloride occurs if these compounds are refluxed together for approximately 24 h. This observation led us to develop a method for the preparation of radioactive phosphorus oxychloride on a small scale with high specific radioactivity. The labeled phosphorus oxychloride may be utilized directly in ...

Progesterone receptor from an oviduct tissue mince incubated with (32P)orthophosphate was purified using affinity chromatography, gel filtration, and DEAE-Sephadex chromatography. Two receptor peaks were eluted from the DEAE column. Peak I contained one major protein band with a molecular weight of 90,000 and a 32P band at 90,000. Peak II also had a major 90,000-dalton protein band and a 32P ba...

A simple, rapid and inexpensive method is described for the enzymic synthesis of [alpha-32P]ATP from [32P]Pi on a preparative scale with an overall yield of 53%. The final product contained all of the detectable radioactivity (less than 99.9%) in the alpha position and has been shown to behave identically with commerically availabe [alpha-32P]ATP during the synthesis of 3':5'-cyclic AMP in the ...

Analysis of aphthovirus A12, strain 119ab, grown in the presence of inorganic 32P revealed that two of the major viral polypeptides, VP4 and trypsin-sensitive protein VP3, were highly phosphorylated. The other major polypeptides, VP1 and VP2, were also phosphorylated but to a much lesser extent. Polypeptides VP0 and P56, of which there are approximately one of two copies per aphthovirion, were ...

Our laboratory has developed a novel delivery platform using an attenuated non-toxic and non-pathogenic bacterium Listeria monocytogenes that infects tumor cells and selectively survives and multiplies in metastases and primary tumors with help of myeloid-derived suppressor cells (MDSC) and immune suppression in the tumor microenvironment (TME). 32P was efficiently incorporated into the Listeri...