Haemoglobin, the`honorary enzyme ' [Brunori (1999), Trends Biochem. Sci. 24, 158±161], constitutes a prime prototype for allosteric models. Here, the crystallization and preliminary X-ray analysis of haemoglobin I from the South American ®sh Brycon cephalus are reported. X-ray diffraction data have been collected to 2.5 A Ê resolution using synchrotron radiation (LNLS). Crystals were determined to belong to the space group P6 1 22 and preliminary structural analysis revealed the presence of one dimer () in the asymmetric unit. The structure was determined using standard molecular-replacement techniques.
The present work reports our succesfull experience concerning crystallization of four fish hemoglobins from three Brazilian species of Teleosts: Liposarcus anisitsi, Brycon cephalus and Piaractus mesopotamicus. The data shown here is part of a systematic functional and structural study of fish hemoglobins with the aim of better understanding the outstanding range of functional and structural properties exhibited by these proteins. We also present a reduced sparse-matrix method for crystallization of fish hemoglobins, which can reduce the amount of hemoglobin initially used in the crystallization experiments.
Haemoglobins constitute a set of proteins with interesting structural and functional properties, especially when the two large animal groups reptiles and fishes are focused on. Here, the crystallization and preliminary X-ray analysis of haemoglobin-II from the South American fish matrinxã (Brycon cephalus) is reported. X-ray diffraction data have been collected to 3.0 A resolution using synchrotron radiation (LNLS). Crystals were determined to belong to space group P2(1) and preliminary structural analysis revealed the presence of two tetramers in the asymmetric unit. The structure was determined using the standard molecular-replacement technique.
Abstract:In this work the authors evaluated the expression of cat, glu, cypP450, hsp70 and 18S RNA in Colossoma macropomum following exposure to copper and cadmium by semi-quantitative RT-PCR and quantitative qRT-PCR. In RT-PCR and qRT-PCR hsp70 expression was highest after exposure to copper for 3 hours (h). glu expression increased after exposure to cadmium, regardless of time. cat expression increased after the first hour of exposure to both metals, and decreased after 3 hs. cyp450 expression was higher when compared to controls in exposed fish regardless of metal and exposure time. cyp450 expression after exposure to copper for 3 h was twice that recorded after 1 h. In the presence of cadmium, cyp450 expression was higher in the animals exposed for 1 h and lower at 3 h. Therefore, toxicology tests and biomonitoring based on molecular markers should focus on a set of molecular and/or environmental parameters, rather than isolated markers.
Analysis of bacterial diversity in soils along the banks of the Solimões and
Negro rivers, state of Amazonas, Brazil, was by partial sequencing of the genes codifying
the rDNA16S region. Diversity of operational taxonomic units (OTU) and of the divergent
sequences obtained were applied in comparative analysis of microbiological diversity in
the two environments, based on richness estimators and OTU diversity indices. The higher
OTU diversity in the Solimões was based on the higher number of parameters that
evoke this. The interaction between the nucleotide sequences of bacteria inhabiting the
two riverine environments indicated that the two microrganism communities are similar in
composition.
Carboxyhaemoglobin-II isolated from the pacu (Piaractus mesopotamicus) has been crystallized and X-ray diffraction data were collected to 2.0 A resolution using synchrotron radiation. Crystals were characterized as belonging to the space group I23; preliminary structural analysis reveals the presence of one dimer in the asymmetric unit.
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