Fig. 6. The kinase activity of Csk is decreased in HIV-infected patients. (A) Csk protein tyrosine kinase was immunoprecipitated from lysates containing equal total protein as measured by the bicinchoninic acid method, from uninfected individuals and from asymptomatic HIV+ patients. The immune complexes were washed, incubated with c-Src C-terminal peptide [Thr-Ser-Thr-Glu-Pro-Gln-Tyr-Gln-Pro-Gly-Glu-Asn-Leu] and 10 μCi [γ32P]ATP at 37°C for 15 minutes, centrifuged, and the supernatant blotted onto phosphocellulose paper and washed with 10% phosphoric acid. After drying, the blotted phosphocellulose paper was tested for 32P-containing c-Src peptide by scintillation counting. The results represent the relative Csk kinase activity of uninfected control and HIV+ patient after subtracting the background control (immune-complex without lysate; 350 cpm). (B) Western immunoblotting of 50-kD Csk protein for (A). Antibody control consists of immune complex without lysate. These results are representative of three independent experiments.
Fig. 6.

The kinase activity of Csk is decreased in HIV-infected patients. (A) Csk protein tyrosine kinase was immunoprecipitated from lysates containing equal total protein as measured by the bicinchoninic acid method, from uninfected individuals and from asymptomatic HIV+ patients. The immune complexes were washed, incubated with c-Src C-terminal peptide [Thr-Ser-Thr-Glu-Pro-Gln-Tyr-Gln-Pro-Gly-Glu-Asn-Leu] and 10 μCi [γ32P]ATP at 37°C for 15 minutes, centrifuged, and the supernatant blotted onto phosphocellulose paper and washed with 10% phosphoric acid. After drying, the blotted phosphocellulose paper was tested for 32P-containing c-Src peptide by scintillation counting. The results represent the relative Csk kinase activity of uninfected control and HIV+ patient after subtracting the background control (immune-complex without lysate; 350 cpm). (B) Western immunoblotting of 50-kD Csk protein for (A). Antibody control consists of immune complex without lysate. These results are representative of three independent experiments.

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