The First Finding of the Actinia Synandwakia hozawai in the Sea of Okhotsk

The finding of the actinia Synandwakia hozawai in the coastal waters of northern Sakhalin (Sea of Okhotsk) suggests a wider range of this species, which was previously only known to inhabit the coastal waters of eastern Japan. Data are presented on the morphology of the S. hozawai specimen from the Sea of Okhotsk and the types of its nematocysts.     

An oligonucleotide microarray for detection and discrimination of orthopoxviruses based on oligonucleotide sequences of two viral genes

An oligonucleotide microarray for detection and identification of orthopoxviruses was developed. Genus specific and orthopoxvirus species-specific regions of the genes encoding chemokine binding and alpha/beta-interferon binding proteins were used as a target. The developed microarray allows the variola, monkeypox, cowpox, vaccinia, camel-pox and ectromelia (mousepox) viruses to be distinguished with a high degree of reliability.     

Activity of caudal mesenteric ganglion in relation to its impulsation (age aspect)

It has been shown in acute experiments on cats and kittens that the pre- and postganglionic nerves of the caudal mesenteric sympathetic ganglion (CMSG) are bioelectrically active from the first day of the animal's life. When the impulsation which comes to the ganglion along the afferent and preganglionic fibers is removed, the impulse activity of the post-ganglionic neurons disappears completely. In newborn kittens the level of activity of the postganglionic fibers is determined to a greater degree by the afferent impulsation of the hypogastric nerves than in adult cats. The effect of the preganglionic impulsation, especially that coming through the intramesenteric nerves, predominates in adult animals. The efferent impulsation of the hypogastric nerves in cats not older than five days disappeared after sectioning of two to three preganglionic branches of the CMSG; in six to nine and 14 to 15-day-old cats, after sectioning of four and in month old cats after sectioning of five; and in two to three month old cats after sectioning of six branches. In adult cats after the first sectioning the postganglionic impulsation frequently intensified, and after sectioning of seven branches the activity completely disappeared. Automatic activity of the ganglia was not observed, postganglionic impulsation developed and changed under the influence of the impulses coming to the ganglion.Scientific Research Institute of Childhood and Adolescent Physiology, Academy of Pedagogical Sciences USSR, Moscow. Translated from Neirofiziologiya, Vol. 3, No. 5, pp. 533–542, September–October, 1971.     

Purification of a recombinant membrane protein tagged with a calmodulin-binding domain: properties of chimeras of the Escherichia coli nicotinamide nucleotide transhydrogenase and the C-terminus of human plasma membrane Ca2+ -ATPase

A Ca2+ -dependent calmodulin-binding peptide (CBP) is an attractive tag for affinity purification of recombinant proteins, especially membrane proteins, since elution is simply accomplished by removing/chelating Ca2+. To develop a single-step calmodulin/CBP-dependent purification procedure for Escherichia coli nicotinamide nucleotide transhydrogenase, a 49 amino acid large CBP or a larger 149 amino acid C-terminal fragment of human plasma membrane Ca2+ -ATPase (hPMCA) was fused C-terminally to the beta subunit of transhydrogenase. Fusion using the 49 amino acid fragment resulted in a dramatic loss of transhydrogenase expression while fusion with the 149 amino acid fragment gave a satisfactory expression. This chimeric protein was purified by affinity chromatography on calmodulin-Sepharose with mild elution with EDTA. The purity and activity were comparable to those obtained with His-tagged transhydrogenase and showed an increased stability. CBP-tagged transhydrogenase contained a 4- to 10-fold higher amount of the alpha subunit relative to the beta subunit as compared to wild-type transhydrogenase. To determine whether the latter was due to the CBP tag, a double-tagged transhydrogenase with both an N-terminal 6x His-tag and a CBP-tag, purified by using either tag, gave no significant increase in purity as compared to the single-tagged protein. The reasons for the altered subunit composition are discussed. The results suggest that, depending on the construct, the CBP-tag may be a suitable affinity purification tag for membrane proteins in general.     

Sequence analysis and expression profiling of 14-3-3 genes from the extremophile <Emphasis Type=Italic>Thelungiella salsuginea</Emphasis>, ecotype Yakutsk

Members of the 14-3-3 protein family are known to be important regulators of plant primary metabolism, hormonal signal transduction, and ion homeostasis. We identified nine isoforms of 14-3-3 genes of Thellungiella salsuginea, an extremophile relative of Arabidopsis thaliana. All the identified isoforms were designated according to their Arabidopsis orthologs: Chi, Omega, Psi, Phi, Upsilon, Lambda, Mu, Epsilon, and Omicron. Comparison of the deduced amino acid sequences reveals high degree of identity between the members of this protein family. Isoforms, designated as Ts14-3-3 Chi, Omicron, and Mu, display noticeable differences in their C-terminal domain as compared to their Arabidopsis homologs. Phylogenetic analysis demonstrated that the identified isoforms split into two groups, epsilon and non-epsilon, according to the common classification of the 14-3-3 family genes. The Thellungiella 14-3-3 isoforms are differentially expressed in various plant tissues, and real-time RT-PCR revealed that most of the isoforms are highly expressed even under normal growth conditions. In response to abiotic stress, low temperatures and high concentrations of salts, 14-3-3 genes exhibited different expression patterns. Our data suggest that, due to the high expression levels of the 14-3-3 genes, Thellungiella plants are likely pre-adapted to the stress conditions. Differences between the C-terminal domains of some Thellungiella 14-3-3 proteins and their Arabidopsis homologs may result in differences in target protein specificity.     

Epitope mapping of monoclonal antibodies to human plasma membrane Ca2-ATPase

Overlapping fragments of the fourth isoform of human plasma membrane Ca(2+)-ATPase (hPMCA4) and several fragments of hPMCA1 were expressed in bacterial cells and purified by metal affinity chromatography. Enzyme immunoassays of the fragments helped map epitopes for 4 monoclonal antibodies (2D8, 8B8, 7C8 and 5E6). The epitope for 2D8 was localized within the 222-249 site (i.e., in the putative transduction domain), the epitopes for 8B8 and 7C8 were localized within the 330-353 site, in which phospholipids are presumably bound, and the 5E6 epitope was found within the 791-843 site, where the putative hinge region is situated. 2D8 recognizes hPMCA1 and hPMCA4 isoforms, while 8B8 and 7C8 are specific for hPMCA4. The amino acid sequences of these epitopes and phage-displayed mimotopes were compared.