M2 pyruvate kinase provides a mechanism for nutrient sensing and regulation of cell proliferation

We show that the M2 isoform of pyruvate kinase (M2PYK) exists in equilibrium between monomers and tetramers regulated by allosteric binding of naturally occurring small-molecule metabolites. Phenylalanine stabilizes an inactive T-state tetrameric conformer and inhibits M2PYK with an IC(50) value of 0.24 mM, whereas thyroid hormone (triiodo-l-thyronine, T3) stabilizes an inactive monomeric form of M2PYK with an IC(50) of 78 nM. The allosteric activator fructose-1,6-bisphosphate [F16BP, AC(50) (concentration that gives 50% activation) of 7 μM] shifts the equilibrium to the tetrameric active R-state, which has a similar activity to that of the constitutively fully active isoform M1PYK. Proliferation assays using HCT-116 cells showed that addition of inhibitors phenylalanine and T3 both increased cell proliferation, whereas addition of the activator F16BP reduced proliferation. F16BP abrogates the inhibitory effect of both phenylalanine and T3, highlighting a dominant role of M2PYK allosteric activation in the regulation of cancer proliferation. X-ray structures show constitutively fully active M1PYK and F16BP-bound M2PYK in an R-state conformation with a lysine at the dimer-interface acting as a peg in a hole, locking the active tetramer conformation. Binding of phenylalanine in an allosteric pocket induces a 13° rotation of the protomers, destroying the peg-in-hole R-state interface. This distinct T-state tetramer is stabilized by flipped out Trp/Arg side chains that stack across the dimer interface. X-ray structures and biophysical binding data of M2PYK complexes explain how, at a molecular level, fluctuations in concentrations of amino acids, thyroid hormone, and glucose metabolites switch M2PYK on and off to provide the cell with a nutrient sensing and growth signaling mechanism

Engineering design of vertical test stand cryostat

Under Indian Institutions and Fermilab collaboration, Raja Ramanna Centre for Advanced Technology and Fermi National Accelerator Laboratory are jointly developing 2K Vertical Test Stand (VTS) cryostats for testing SCRF cavities at 2K. The VTS cryostat has been designed for a large testing aperture of 86.36 cm for testing of 325 MHz Spoke resonators, 650 MHz and 1.3 GHz multi-cell SCRF cavities for Fermilab's Project-X. Units will be installed at Fermilab and RRCAT and used to test cavities for Project-X. A VTS cryostat comprises of liquid helium (LHe) vessel with internal magnetic shield, top insert plate equipped with cavity support stand and radiation shield, liquid nitrogen (LN{sub 2}) shield and vacuum vessel with external magnetic shield. The engineering design and analysis of VTS cryostat has been carried out using ASME B&PV Code and Finite Element Analysis. Design of internal and external magnetic shields was performed to limit the magnetic field inside LHe vessel at the cavity surface <1 {micro}T. Thermal analysis for LN{sub 2} shield has been performed to check the effectiveness of LN{sub 2} cooling and for compliance with ASME piping code allowable stresses

Vermiculture Biotechnology: The Emerging Cost-Effective and Sustainable Technology of the 21st Century for Waste and Land Management to Safe and Sustainable Food Production

Griffith Sciences, Griffith School of EngineeringNo Full Tex