The regulation of protein kinase C by thiol oxidation

The Ser/Thr protein kinase C (PKC) isozyme family plays an important role in cell growth and differentiation and also contributes to key events in the development and progression of cancer. PKC isozymes are activated by phospholipid-dependent mechanisms, and they are also subject to oxidative activation and inactivation. Oxidative regulatory mechanisms are important in the governance of PKC isozyme action. While oxidative PKC activation involves phospho-tyrosine (P-Y) stabilization, the molecular mechanism(s) for oxidative PKC inactivation have not been defined. We previously reported that Thr → Cys peptide-substrate analogs inactivate several PKC isozymes including PKC-α via S-thiolation, i.e., by forming disulfides with PKC thiols. This inactivation mechanism is chemically analogous to protein S-glutathiolation, a post-translational modification that has been shown to oxidatively regulate several enzymes. To determine if PKC-α could be inactivated by S-glutathiolation, we employed the thiol-specific oxidant diamide (0.01–10mM) and 100μM glutathione (GSH). Diamide alone (0.1–5.0 mM) weakly inactivated PKC-α (<20%), and GSH alone had no effect on the isozyme activity. Marked potentiation of diamide-induced PKC-α inactivation (>90%) was achieved by 100μM GSH, resulting in full inactivation of the isozyme. Inactivation was reversed by DTT, consistent with a mechanism involving PKC-α S-glutathiolation. S-glutathiolation was demonstrated as DTT-reversible incorporation of [35S] GSH into PKC-α isozyme structure. These results indicate that a mild oxidative stimulus can inactivate purified PKC-α via S-glutathiolation. In addition, diamide treatment of metabolically labeled NIH3T3 cells induced potent PKC-α inactivation via isozyme [35S] S-thiolation. These results indicate that cellular PKC-α can be regulated via S-glutathiolation. ^

Manzano : : distribución de micronutrientes en suelo

Para medir la disponibilidad de los micronutrientes en la zona de mayor densidad radical de los frutales de pepita y carozo y sugerir pautas de manejo que permitan un uso sustentable del recurso suelo, se tomaron muestras representativas de 25 montes cultivados con manzanos a lo largo del Alto Valle del río Negro (Argentina). Se extrajeron muestras a 0-25 y 25-50 cm de profundidad y se determinó la concentración disponible de Fe, Cu, Mn y Zn; granulometría; pH; materia orgánica (MO); carbonatos; P y capacidad de intercambio catiónico (CIC). Los resultados muestran que los micronutrientes se concentran mayoritariamente en la primera capa de suelo analizada, disminuyendo abruptamente en el estrato 25-50 cm. En la capa superficial, la disponibilidad de Cu y Zn está influenciada por el P mientras que el pH afecta la del Fe, Cu, y Mn en el estrato de suelo de 25-50 cm. En conclusión, es posible mejorar la nutrición mineral de los cultivos si se crean condiciones favorables para el crecimiento radical en la capa superficial del suelo.