Structure-function relationship of new crotamine isoform from the Crotalus durissus cascavella

In this work we isolated a novel crotamine like protein from the Crotalus durissus cascavella venom by combination of molecular exclusion and analytical reverse phase HPLC. Its primary structure was:YKRCHKKGGHCFPKEKICLPPSSDLGKMDCRWKRK-CCKKGS GK. This protein showed a molecular mass of 4892.89 da that was determined by Matrix Assisted Laser Desorption Ionization Time-of-flight (MALDI-TOF) mass spectrometry. The approximately pI value of this protein was determined in 9.9 by two-dimensional electrophoresis. This crotamine-like protein isolated here and that named as Cro 2 produced skeletal muscle spasm and spastic paralysis in mice similarly to other crotamines like proteins. Cro 2 did not modify the insulin secretion at low glucose concentration (2.8 and 5.6 mM), but at high glucose concentration (16.7 mM) we observed an insulin secretion increasing of 2.7-3.0-fold than to control. The Na+ channel antagonist tetrodoxin (6 mM) decreased glucose and Cro 2-induced insulin secretion. These results suggested that Na+ channel are involved in the insulin secretion. In this article, we also purified some peptide fragment from the treatment of reduced and carboxymethylated Cro 2 (RC-Cro 2) with cyanogen bromide and protease V8 from Staphylococcus aureus. The isolated pancreatic beta-cells were then treated with peptides only at high glucose concentration (16.7 mM), in this condition only two peptides induced insulin secretion. The amino acid sequence homology analysis of the whole crotamine as well as the biologically-active peptide allowed determining the consensus region of the biologically-active crotamine responsible for insulin secretion was KGGHCFPKE and DCRWKWKCCKKGSG.

Burrows with Chimneys of the Fiddler Crab Uca thayeri: Construction, Occurrence, and Function

Joao B. L. Gusmao-Junior, Glauco B. O. Machado, and Tania M. Costa (2012) Burrows with chimneys of the fiddler crab Uca thayeri: construction, occurrence, and function. Zoological Studies 51(5): 598-605. Building of soil structures is observed in a variety of semi-terrestrial crustaceans. In fiddler crabs (Genus Uca), this behavior occurs in several species, some of which build structures that are largely ornamental and others construct barriers that are apparently for defense. Although there is a relative abundance of studies on this type of behavior in Uca, the relationship between the social context and the occurrence of these structures remains poorly studied. Thus, this study attempted to analyze in detail the construction, occurrence, and function of mud chimneys built by the fiddler crab Uca thayeri; these sedimentary structures are possibly associated with burrow defense. Field investigations and laboratory experiments were conducted. Both sexes were often found in burrows with chimneys; however, laboratory experiments showed that only females actively built and maintained chimneys, with some difference in the morphology of these structures between sexes. The social context had little influence on the construction of chimneys, which showed that the stimulus for constructing chimneys could be endogenous. Our results suggest that burrows with chimney of U. thayeri may have functions other than defense, and may act in regulating the internal conditions of the burrow, as observed in other crustaceans with such building behavior. http://zoolstud.sinica.edu.tw/Journals/51.5/598.pdf