Site-specific N-Linked Glycosylation of Receptor Guanylyl Cyclase C Regulates Ligand Binding, Ligand-mediated Activation and Interaction with Vesicular Integral Membrane Protein 36, VIP36

Guanylyl cyclase C (GC-C) is a multidomain, membrane-associated receptor guanylyl cyclase. GC-C is primarily expressed in the gastrointestinal tract, where it mediates fluid-ion homeostasis, intestinal inflammation, and cell proliferation in a cGMP-dependent manner, following activation by its ligands guanylin, uroguanylin, or the heat-stable enterotoxin peptide (ST). GC-C is also expressed in neurons, where it plays a role in satiation and attention deficiency/hyperactive behavior. GC-C is glycosylated in the extracellular domain, and differentially glycosylated forms that are resident in the endoplasmic reticulum (130 kDa) and the plasma membrane (145 kDa) bind the ST peptide with equal affinity. When glycosylation of human GC-C was prevented, either by pharmacological intervention or by mutation of all of the 10 predicted glycosylation sites, ST binding and surface localization was abolished. Systematic mutagenesis of each of the 10 sites of glycosylation in GC-C, either singly or in combination, identified two sites that were critical for ligand binding and two that regulated ST-mediated activation. We also show that GC-C is the first identified receptor client of the lectin chaperone vesicular integral membrane protein, VIP36. Interaction with VIP36 is dependent on glycosylation at the same sites that allow GC-C to fold and bind ligand. Because glycosylation of proteins is altered in many diseases and in a tissue-dependent manner, the activity and/or glycan-mediated interactions of GC-C may have a crucial role to play in its functions in different cell types.

Multidimensional free energy surface of unfolding of HP-36: Microscopic origin of ruggedness

The protein folding funnel paradigm suggests that folding and unfolding proceed as directed diffusion in a multidimensional free energy surface where a multitude of pathways can be traversed during the protein's sojourn from initial to final state. However, finding even a single pathway, with the detail chronicling of intermediates, is an arduous task. In this work we explore the free energy surface of unfolding pathway through umbrella sampling, for a small globular a-helical protein chicken-villin headpiece (HP-36) when the melting of secondary structures is induced by adding DMSO in aqueous solution. We find that the unfolding proceeds through the initial separation or melting of aggregated hydrophobic core that comprises of three phenylalanine residues (Phe7, Phe11, and Phe18). This separation is accompanied by simultaneous melting of the second helix. Unfolding is found to be a multistage process involving crossing of three consecutive minima and two barriers at the initial stage. At a molecular level, Phe18 is observed to reorient itself towards other hydrophobic grooves to stabilize the intermediate states. We identify the configuration of the intermediates and correlate the intermediates with those obtained in our previous works. We also give an estimate of the barriers for different transition states and observe the softening of the barriers with increasing DMSO concentration. We show that higher concentration of DMSO tunes the unfolding pathway by destabilizing the third minimum and stabilizing the second one, indicating the development of a solvent modified, less rugged pathway. The prime outcome of this work is the demonstration that mixed solvents can profoundly transform the nature of the energy landscape and induce unfolding via a modified route. A successful application of Kramer's rate equation correlating the free energy simulation results shows faster rate of unfolding with increasing DMSO concentration. This work perhaps presents the first systematic theoretical study of the effect of a chemical denaturant on the microscopic free energy surface and rates of unfolding of HP-36. (C) 2014 AIP Publishing LLC.

Prudentia Iuris, 1994, n° 36 (número completo)

Contenido: La inserción de la universidad en el medio : país e Iglesia / Domingo Basso – Reflexiones en torno al ser y al hacer de las universidades católicas / Jorge Humberto Peláez – La Constitución y los tratados internacionales a partir de la reforma de 1994 / Eugenio Luis Palazzo – Los Iudicia Bonae Fidei y su importancia en materia contractual / Alberto Gustavo Di Pietro – En torno a Pro Caccina / Edmundo J. Carbone – Los iuris praecepta o la norma ética como imperativo jurídico / Ángel Hugo Guerriero – La objetividad en el saber político / Héctor Julio Martinotti – Presencia de la Iglesia en la universidad y en la cultura universitaria / Pío Laghi ; Eduardo Pironio ; Paul Poupard – III Encuentro Internacional de Derecho de América del Sur. “El derecho y la integración hacia el siglo XXI”. “Pasos previos para la integración política de América Latina” / Flavio Fernando Agatiello Piñero

Modelo de solicitud de documentación sobre la Guerra Civil del 36 elaborado por el Centro Vasco de Caracas.

Fecha: >1970 / Unidad de instalación: Carpeta 48 - Expediente 7-14 / Nº de pág.: 3 (mecanografiadas)

Aquaculture Asia, Vol. 11, No. 1, pp.1-36, January - March 2006

*Table of Contents* Research & farming techniques Nursery rearing of Puntius goniotus: A preliminary trial K.N. Mohnta, J.K. Jena & S.N. Mohanty Artemia enrichment and biomass production for larval finfish and shellfish culture A.S. Ninawe Vembanad Lake: A potential spawner bank of the giant freshwater prawn Macrobrachium rosenbergii on the southwest coast of India Paramaraj Balamurugan, Pitchaimuthu Mariappan & Chellam Balasundaram Seed production of mud crab Scylla serrata at the Rajiv Gandhi Center for Aquaculture, Tamil Nadu, India Mohamed Shaji, Emilia T. Quinitio, Thampi Samraj, S. Kandan, K. Ganesh, Dinesh Kumar, S. Arulraj, S. Pandiarajan, Shajina Ismail and K. Dhandapan. Sustainable aquaculture Fish wastes in urban and suburban markets of Kolkata: Problems and potentials Kausik Mondal, Anilava Kaviraj & P.K. Mukhopadhyay People in aquaculture Peter Edwards writes on rural aquaculture: Farming carps in leased ponds by groups of poor women in Chandpur, Bangladesh Aquatic animal health Lymphocystis disease and diagnostic methods in China Jing Xing, Xiuzhen Sheng & Wenbin Zhan Asia-Pacific Marine Finfish Aquaculture Network Mesocosm technology advances grouper culture in northern Australia Elizabeth Cox, Peter Fry & Anjanette Johnston

The diatoms of Longans lake-vegetation. Fossil diatom flora. [Translation from: Acta Phytogeographica Suecica 36 73-86, 1955. ]

Sediment samples were taken from Lake Langans in Sweden and fossilised diatoms analysed. Sample methods and environmental factors are discussed. Species with a characteristic occurrence are described. The article discusses diatom-thanatocoenoses as indicators of environment.

On the problem of the distribution of Acanthocyclops viridis Jurine in the Rybinsk reservoir. [Translation from: Byulleten' Institute Biologii Vodokhranilishch 1958(1), 36-39, 1958. ]

Observations of individual weight, duration of development and production of different stages of Tropodiaptomus incognitus are presented. The study is based on data gathered from Lake Chad in 1968.

Studies on the mouth size of fish larvae. Method and conclusions only [Translation from: Bulletin of the Japanese Society of Scientific Fisheries 36, 353-368, 1970]

This study was conducted to look into the relationship between mouth size and the total length of larval fish, and the growth in total length of larval fish in one or within a one-year period. Study material was gathered in the South China Sea, the Bay of Bangkok and the Vietnamese coast. This translation focuses on methods and conclusions of the original (longer) paper.

Influence of carbon tetrachloride on the liver of tadpoles of brown frog (Rana temporaria) [Translation from: Trudy leningt.sanit.-gig.med.Inst. 82, 32-36, 1967]

Comparative studies of the structure and regeneration of tissues in different vertebrates make it possible to get ideas from the evolution and regeneration potential of tissues. In this study are considered the reactive changes in the liver of larvae of the amphibian Rana temporaria to CC14 of different concentrations. Tadpoles of different sizes and different growth were placed once each day every day in a vessel with a determined solution of CC14 in water. The liver of tadpoles has a distinctive structure. It is distinguished by structure not only from the liver of higher vertebrates, but also from the liver of the adult frogs. The liver of the latter has an insignificant amount of fat and a whole series of other characters not typical of the liver of the tadpole. Placing the tadpoles in a solution of CCl4 with a concentration of 0.1% did not produce in the liver noticeable morphological changes. The author concludes that the absence of degenerative changes in the liver tadpoles, in spite of a high percentage of death of the experimental animals, tells of the well-known resistance of their liver to the influence of CC14.

Some data on the content of organic carbon in rivers in the winter months. [Translation from: Gidrochim. Mat. 23, 36-38, 1955]

Observations are reported on the content of organic matter in the Moscow region in 1941. Some data is given on alkalinity, oxygen content and colourisation of the rivers.