Fusion polypeptides in gene cloning: potential problems due to conformational alterations at the junction

Many eukaryotic genes are cloned in bacterial hosts as fusion polypeptides. Prediction of the secondary structures for some common prokaryotic fusion polypeptides shows that many junction sites correspond to important secondary structures. It is suggested that such structures could affect (hinder, etc.) the conformation or drive the folding of the neighboring eukaryotic counterparts. Thus the prokaryotic junction should be better performed in random coil regions, or short fusion prokaryotic polypeptides should be used.     

Occludin binds to the SH3-hinge-GuK unit of zonula occludens protein 1: potential mechanism of tight junction regulation

The interaction between tight junction proteins occludin and zona occludens protein 1 (ZO-1) was clarified. The sequence cc1 within the hinge region of ZO-1, connecting its SH3 and GuK domains, was identified as a new association site for the occludin C-terminus, core binding area GLRSSKRNLRKSR (mouse ZO-1^606-618). Occludin also bound to the sequence H2 within GuK, core area HKLRKNNH (ZO-1^759-766). In occludin, the binding core was ELSRLDKELDDYREESEEY (mouse occludin^455-473). Helicity of the sequences was suggested by circular dichroism. Because basic residues in ZO-1, acidic residues in occludin (underlined), coiled-coil helix-forming leucine heptad motifs (bold) in occludin and, probably, in cc1 were essential, we conclude that interactions were both helical and ionic. Moreover, the GuK domain bound other GuK molecules, suggesting oligomerization of ZO-1. Generally, the assumption is supported that the SH3-hinge-GuK region represents a functional and regulatory unit in ZO-1 forming a multiprotein tight junction complex with occludin.Received 12 January 2004; received after revision 23 February 2004; accepted 31 March 2004     

Volatile compounds associated with estrus in mouse urine: potential pheromones

Female mice that had been made estrous through hormone implantation excreted in their urine significantly enhanced levels of n-pentyl acetate, cis-2-penten-1-yl acetate, p-toluidine, 2-heptanone, and 3 unsaturated ketones. The relationship of these volatiles to a signaling function of the estrous urine is postulated. Structural elucidations of these compounds were carried out through capillary gas chromatography/mass spectrometry and the synthesis of authentic samples.     

Afferent nerve endings in the avian lung: Observations with the light microscope

Résumé Des examens au microscope optique on montré l'existence de terminaisons nerveuses, dont on supposait la présence, dans les structures des bronchioles tertiaires et des atrium des poumons de la poule domestique (G. domesticus). Les caractères morphologiques en sont décrits. On pense qul'il s'agit de la première preuve de l'existence de terminaisons afférentes dans les «conduits» d'air, réalisée à l'aide du microscope optique.     

A new stimulation technique of the crista ampullaris of the lateral canal in the adult cat: Study of the action potential of the vestibular nerve

Summary The stimulation of the crista ampullaris of the lateral canal by a short ampullopetal liquid flux produces on the pre-ganglionic vestibular fibres the appearance of a bimodal action potential. The amplitude of this action potential increases with the intensity of the stimulation. This stimulation also provokes the birth of an evoked potential at the level of the vestibular nuclei as well as an ocular jerk.Supported by the D.G.R.S.T. grant 77.7.0669 and I.N.S.E.R.M. (A.S.R. No. 6, grant 011).     

A new stimulation technique of the crista ampullaris of the lateral canal in the adult cat: study of the action potential of the vestibular nerve.

The stimulation of the crista ampullaris of the lateral canal by a short ampullopetal liquid flux produces on the pre-ganglionic vestibular fibres the appearance of a bimodal action potential. The amplitude of this action potential increases with the intensity of the stimulation. This stimulation also provokes the birth of an evoked potential at the level of the vestibular nuclei as well as an ocular jerk.