A Monoclonal Antibody that Specifically Recognizes m'A Nucleoside

Abstract

A hybridoma against the nucleoside m⁶A has been obtained from mouse spleen. This hybridoma was named H65 and it secretes monoclonal antibodies anti-m⁶A. The competition assays showed that the monoclonal antibody was highly specific for m⁶A nucleoside.     

Structure-Function Features of a Mycoplasma Glycolipid Synthase Derived from Structural Data Integration,Molecular Simulations,and Mutational Analysis

Glycoglycerolipids are structural components of mycoplasma membranes with a fundamental role in membrane properties and stability. Their biosynthesis is mediated by glycosyltransferases (GT) that catalyze the transfer of glycosyl units from a sugar nucleotide donor to diacylglycerol. The essential function of glycolipid synthases in mycoplasma viability, and the absence of glycoglycerolipids in animal host cells make these GT enzymes a target for drug discovery by designing specific inhibitors. However, rational drug design has been hampered by the lack of structural information for any mycoplasma GT. Most of the annotated GTs in pathogenic mycoplasmas belong to family GT2. We had previously shown that MG517 in Mycoplasma genitalium is a GT-A family GT2 membrane-associated glycolipid synthase. We present here a series of structural models of MG517 obtained by homology modeling following a multiple-template approach. The models have been validated by mutational analysis and refined by long scale molecular dynamics simulations. Based on the models, key structure-function relationships have been identified: The N-terminal GT domain has a GT-A topology that includes a non-conserved variable region involved in acceptor substrate binding. Glu193 is proposed as the catalytic base in the GT mechanism, and Asp40, Tyr126, Tyr169, Ile170 and Tyr218 define the substrates binding site. Mutation Y169F increases the enzyme activity and significantly alters the processivity (or sequential transferase activity) of the enzyme. This is the first structural model of a GT-A glycoglycerolipid synthase and provides preliminary insights into structure and function relationships in this family of enzymes.     

Life history and production of the burrowing mayfly Ephoron virgo (Olivier, 1791) (Ephemeroptera: Polymitarcyidae) in the lower Ebro river: a comparison after 18 years

The life history of the burrowing mayfly Ephoron virgo (Olivier, 1791) (Ephemeroptera: Polymitarcyidae) was studied during spring and summer 2005 in the lower Ebro river (Catalonia) and compared to a previous study performed in 1987 (Ibáñez, Escosa, Muñoz and Prat 1991). The results showed an advancement of Ephoron virgo life cycle and an increase of production estimates. In 2005 larval development reached the maximum size one month earlier than in 1987, and adult emergence peak began three weeks earlier. Comparing adult sex ratios (F:M), there was a major presence of females in 2005 (1:4), while the opposite was observed in 1987 (2:1). Secondary production was higher in 2005 than in 1987, obtaining 950 mg dry weight/m²/year with the increment summation method and 1080 mg dry weight/m²/year using the removal summation method. Higher water temperatures were measured for the entire 2005 larval growth period, which were related to higher air temperatures. Therefore, that temperature increment was likely the main cause of changes observed in the Ephoron virgo life cycle.