An AKAP-Lbc-RhoA interaction inhibitor promotes the translocation of aquaporin-2 to the plasma membrane of renal collecting duct principal cells

Stimulation of renal collecting duct principal cells with antidiuretic hormone (arginine-vasopressin, AVP) results in inhibition of the small GTPase RhoA and the enrichment of the water channel aquaporin-2 (AQP2) in the plasma membrane. The membrane insertion facilitates water reabsorption from primary urine and fine-tuning of body water homeostasis. Rho guanine nucleotide exchange factors (GEFs) interact with RhoA, catalyze the exchange of GDP for GTP and thereby activate the GTPase. However, GEFs involved in the control of AQP2 in renal principal cells are unknown. The A-kinase anchoring protein, AKAP-Lbc, possesses GEF activity, specifically activates RhoA, and is expressed in primary renal inner medullary collecting duct principal (IMCD) cells. Through screening of 18,431 small molecules and synthesis of a focused library around one of the hits, we identified an inhibitor of the interaction of AKAP-Lbc and RhoA. This molecule, Scaff10-8, bound to RhoA, inhibited the AKAP-Lbc-mediated RhoA activation but did not interfere with RhoA activation through other GEFs or activities of other members of the Rho family of small GTPases, Rac1 and Cdc42. Scaff10-8 promoted the redistribution of AQP2 from intracellular vesicles to the periphery of IMCD cells. Thus, our data demonstrate an involvement of AKAP-Lbc-mediated RhoA activation in the control of AQP2 trafficking

Assimilating atmospheric data into a terrestrial biosphere model: A case study of the seasonal cycle

This paper demonstrates a new method of assimilating atmospheric concentration data into terrestrial biosphere models. Using a combination of adjoint and tangent linear models of both the underlying biosphere model and the atmospheric transport model, we directly infer optimal model parameters and their uncertainties. We also compute biospheric fluxes and their uncertainties arising from these parameters. We demonstrate the method using the Simple Diagnostic Biosphere Model (SDBM) and data on the seasonal cycle of CO₂ from 41 observing sites. In the model, the light-use efficiency for several biomes is well-constrained by concentration observations. Optimal values generally increase with latitude as required to match the seasonal cycle. Modeled Q(10) values are poorly constrained unless local flux measurements are also used. Values also increase with latitude but are less than the commonly assumed value of 2