Protein microinjection by protease permeabilization of fibroblasts.

نویسندگان

  • R Lemons
  • S Forster
  • J Thoene
چکیده

Exposure of cultured diploid fibroblasts to protease solutions induces a hyperpermeable state which permits entry of exogenous macromolecules directly into the cytosol. We have exploited this finding to devise a microinjection method whose chief advantages are simplicity and good retention of cell viability. Proteins successfully injected by this technique range from insulin to thyroglobulin. The amounts injected range from 4 X 10(5) to 5 X 10(6) molecules/cell.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Modulation of vimentin containing intermediate filament distribution and phosphorylation in living fibroblasts by the cAMP-dependent protein kinase

Microinjection of the purified catalytic subunit of the cAMP-dependent protein kinase (A-kinase) into living rat embryo fibroblasts leads to dramatic changes in vimentin intermediate filament (IF) organization, involving the collapse of the filaments into tight bundles. In some cell types, this rearrangement of the IF proceeds further, leading to an apparent loss of filament integrity, resultin...

متن کامل

Heat Shock Protein 70 Promotes Cell Survival by Inhibiting Lysosomal Membrane Permeabilization

Heat shock protein 70 (Hsp70) is a potent survival protein whose depletion triggers massive caspase-independent tumor cell death. Here, we show that Hsp70 exerts its prosurvival function by inhibiting lysosomal membrane permeabilization. The cell death induced by Hsp70 depletion was preceded by the release of lysosomal enzymes into the cytosol and inhibited by pharmacological inhibitors of lyso...

متن کامل

Requirement of ATM in phosphorylation of the human p53 protein at serine 15 following DNA double-strand breaks.

Microinjection of the restriction endonuclease HaeIII, which causes DNA double-strand breaks with blunt ends, induces nuclear accumulation of p53 protein in normal and xeroderma pigmentosum (XP) primary fibroblasts. In contrast, this induction of p53 accumulation is not observed in ataxia telangiectasia (AT) fibroblasts. HaeIII-induced p53 protein in normal fibroblasts is phosphorylated at seri...

متن کامل

Effect of thiourea on proteins, catalase, guaiacol-peroxidase and protease activities in wheat leaves under H2O2 induced oxidative stress . Ajmad Hameed1*, Liala Jafri2 and Muneer A. Sheikh2

The effect of different thiourea (TU) concentrations (10 and 20 mM) on antioxidants, protease activity and protein content with and without H2O2 (50 mM) induced stress was investigated in wheat leaves. A gradual decrease in protein content with a peak at 72 hours was observed in stressed as well as control leaves. This gradual decrease in leaf protein content at different time intervals was com...

متن کامل

Degradation of proteins microinjected into IMR-90 human diploid fibroblasts

Erythrocyte ghosts loaded with 125I-labeled proteins were fused with confluent monolayers of IMR-90 fibroblasts using polyethylene glycol. Erythrocyte-mediated microinjection of 125I-proteins did not seriously perturb the metabolism of the recipient fibroblasts as assessed by measurements of rates of protein synthesis, rates of protein degradation, or rates of cellular growth after addition of ...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:
  • Analytical biochemistry

دوره 172 1  شماره 

صفحات  -

تاریخ انتشار 1988