Neurotrophic factor structures reveal clues to evolution, binding, specificity, and receptor activation

The neutrophin family, the glial-derived neurotrophic factor family, and the ciliary neurotrophic factor are the best described growth factors specific for developing neurons and neutral crest cells. As might be expected for regulatory molecules of the complex central and peripheral nervous system, these factors show considerable receptor specificity and cross-talk. Thanks to a decade of intense research by numerous laboratories, the structures of many of these factors are now available. This review discusses the structural bases of receptor binding, specificity, and activation in each of these systems. Using structure-based sequence alignments, the evolutionary implications of these molecules and their receptors are discussed, followed by suggestions for further directions for research on the structure and function of these neurotrophic factors.     

The role of glycosylation in ionotropic glutamate receptor ligand binding, function, and trafficking

Members of the ionotropic glutamate receptor (iGluR) family have between 4 and 12 consensus asparagine (N)-linked glycosylation sites. They are localized on the extracellular N-termini, and the loop between the penultimate and last transmembrane domains. These regions also contain the essential elements for formation of the ligand binding site. N-linked glycosylation does not appear to be essential for formation of the ligand binding site per se, but there are demonstrated interactions between glycosylation state and ligand binding affinity, receptor physiology, susceptibility to allosteric modulation and, in some cases, trafficking. There is no indication of a general role for N-linked glycosylation in iGluRs; instead the effects of glycosylation vary among glutamate receptor subtypes and splice variants, with specific effects on structure or function with different subunits.     

Synthesis and cannabinoid receptor binding activity of conjugated triene anandamide,a novel eicosanoid

A polyenoic fatty-acid isomerase (PFI) from a red marine alga was used to convert anandamide (5Z,8Z,11Z,14Z-eicosatetraenoyl-N-ethanolamide) to the 5Z,7E,9E,14Z-eicosatetraenoyl-N-ethanolamide isomer. This novel eicosanoid, termed conjugated triene anandamide (CTA), was assessed for its ability to bind to the cannabinoid receptor in rat brain membrane preparations. CTA is a high affinity cannabimimetic substance whose novel structure provides new insight into structure-activity relationships of cannabinoid receptor ligands. These experiments illustrate the utility of enzymes isolated from marine organisms in the development of pharmacological probes.     

Identification of human serum proteins binding iron,copper, and thyroid hormones by starch gel electrophoresis

Résumé On a analysé au moyen d'électrophorèse sur gel d'amidon les protéines qui, dans le sérum humain, lient le fer, le cuivre et l'hormone thyroïdienne, en utilisant comme témoins des protéines purifiées et la radioactivité. Le fer est lié par la -globuline qui, dans certaines familles, paraît se diviser en deux composants. Le cuivre est lié par la céruloplasmine, qui émigre dans la position ₂ rapide. La protéine principale liant la thyroxine est la préalbumine.     

In vitro binding of citrinin to serum protein

Summary In vitro study shows that the mycotoxin citrinin binds to human serum albumin.Part of Ph. D. thesis, University of Madras, 1973.Acknowledgment is due to Dr E. R. B. Shanmugasundaram (University of Madras) and Dr J. Jayaraman (Madurai University) for kind guidance.     

TRH analogue with C-terminal thioamide group. Synthesis, receptor binding, TSH-releasing activity and alpha-MSH-releasing activity

A new TRH analogue containing a C-terminal thioamide group was synthesized. This peptide was shown to have receptor-binding affinity, and TSH- as well as alpha-MSH-releasing activities very similar to native TRH.     

Specific in vivo binding of d-LSD in rat brain

Summary A reproducible in vivo d-LSD binding method in rat brain is described, with high affinity (K_d of 5 pmoles/g wet wt), stereospecificity (d-vs, vs. l-LSD) and regional selectivity. It may be a useful adjunct to in vitro methods for measuring changes in turnover at the synaptic level related to the intact receptor.     

Flavodoxins: sequence, folding, binding, function and beyond

Flavodoxins are electron-transfer proteins involved in a variety of photosynthetic and non-photosynthetic reactions in bacteria, whereas, in eukaryotes, a descendant of the flavodoxin gene helps build multidomain proteins. The redox activity of flavodoxin derives from its bound flavin mononucleotide cofactor (FMN), whose intrinsic properties are profoundly modified by the host apoprotein. This review covers the very exciting last decade of flavodoxin research, in which the folding pathway, the structure and stability of the apoprotein, the mechanism of FMN recognition, the interactions that stabilize the functional complex and tailor the redox potentials, and many details of the binding and electron transfer to partner proteins have been revealed. The next decade should witness an even deeper understanding of the flavodoxin molecule and a greater comprehension of its many physiological roles. The fact that flavodoxin is essential for the survival of some human pathogens could make it a drug target on its own. Received 26 October 2005; received after revision 20 November 2005; accepted 14 December 2005     

Specific in vivo binding of d-LSD in rat brain

A reproducible in vivo d-LSD binding method in rat brain is described, with high affinity (Kd of 5 pmoles/g wet wt), stereospecificity (d- vs. 1-LSD) and regional selectivity. It may be a useful adjunct to in vitro methods for measuring changes in turnover at the synaptic level related to the intact receptor.     

FSH receptor binding inhibiting activity associated with low molecular weight inhibin from sheep,human, rat and chicken

Summary Low molecular weight inhibin (1500 daltons) was obtained from sheep, human, rat and chicken testes by sequential chromatography on Sephadex G-100 and G-25. In addition to its ability to suppress circulating FSH levels in adult castrated male rats, it also exhibits binding inhibition of I¹²⁵hFSH to rat testicular receptors.     

Tissue-specific modulation of rat glucocorticoid receptor binding activity by melatonin

The effect of melatonin and 2-Iodomelatonin on nuclear and cytosolic glucocorticoid receptors in the brain, pituitary, thymus and liver has been examined. The results indicate that both melatonin and 2-Iodomelatonin administration is associated with marked changes in the density and the affinity of cytosolic and nuclear forms of glucocorticoid receptors. These observations are discussed in the context of a possible involvement of pineal melatonin in the mechanisms regulating the behaviour and metabolism of steroid receptors.     

Extraction and purification of brain glycoprotein NSA3 by binding with hyaluronic acid

Brain glycoprotein NSA3 was found to bind to immobilised hyaluronic acid. The binding was reversible and gave pure antigen (99%) in high yields (80%). The binding was suppressed by incubating the affinosorbent with hyaluronidase. It was not suppressed by trypsin. The presence of glycosaminoglycans other than hyaluronic acid in the sample did not inhibit the binding. This property is relevant to those already known for the brain glycoprotein NSA3, and to its localisation at the nodes of Ranvier. We propose to coin the name hyaluronectin for the brain glycoprotein NSA3.     

Species-dependent stereospecific serum protein binding of the oral anticoagulant drug phenprocoumon

Summary 13 mammalian species are classified into 3 clearcut groups with respect to the stereospecific serum proteinbinding of phenprocoumon: 2 groups showing opposed stereospecific binding characteristics and a 3rd group exhibiting no stereospecific binding. Structural differences in the albumin molecule account for these stereospecific differences in serum protein-binding.This research was supported by a grant from the Deutsche Forschungsgemeinschaft (Ja/185/6).     

The binding of C14 labelled bacterial polysaccharide to serum proteins

Zusammenfassung Die Bindung zwischen C¹⁴-markiertem Polysaccharid ausK. pneumoniae und Plasmaproteinen von Mensch, Meerschweinchen, Ratte, Maus und Hase wurde papierelektrophoretisch untersucht. Inin vitro-Versuchen konnte gezeigt werden, dass die Bindung zwischen Albumin und C¹⁴-markiertem Polysaccharid bei der Ratte am ausgeprägtesten, beim Meerschweinchen am geringsten ist.In vivo-Versuche ergaben jedoch eine erhebliche Bindung zwischen Albumin und Polysaccharid in beiden Gattungen.     

Regional variations and particle binding of free amino acids in the rat brain: effects of amphetamine and methaqualone]

Amphetamine and methaqualone affected aspartate level in various regions of Rat brain. Increased aspartate level in cerebral cortex of methaqualone treated Rats was produced in the aspartate fraction whose binding to subcellular particules was Na+ dependant. This work suggest that aspartate level was altered only in brain fraction implicated in the pharmacological effects of the drugs.     

Fixation of polyunsaturated fatty acids by alpha fetoprotein and serum albumin in rats. Comparison with the accumulation of these acids in developing rat brain]

Quantitative data are presented on the fatty acid composition of rat alpha-fetoprotein (AFP) and serum albumin, (SA), and of brain extracts of suckling rats. In AFP and SA preparations, 40% and 13%, respectively, of total fatty acids present are polyenoic acids. Among them, docosahexaenoic acid is quantitatively the most important in AFP, while in SA, arachidonic acid is largely predominant. Both docosahexaenoic and arachidonic acids were the predominant polyenoic acids in brain extracts. The rate of accumulation of these acids in the brain of suckling rats and the rate of AFP secretion during the same period showed a maximum around 10--12 days after birth. These results suggest that AFP and SA play an important role in the transport and the incorporation of polyunsaturated fatty acids in the developing brain.