Relationship between gliadin peptide structure and their effect on the fetal chick duodenum

Summary The tendency to form a-turn in-gliadin was estimated using the B-cell determinant prediction program based on the Chou and Fasman probability of-turn formation. Six sequences possessing a high probability of-turn formation were found. A statistically high agreement was found between these six sequences and three areas in-gliadin with the occurrence of Pro-Ser-Gln-Gln sequence which has recently been considered responsible for toxicity in coeliac disease. By means of solid-phase synthesis seven peptides were obtained covering the above-mentioned regions. Their toxicity was tested using the fetal chick duodenum. The results support the suggestion that peptides containing the sequences Pro-Ser-Gln-Gln and Gln-Gln-Gln-Pro may be involved in the pathogenesis of coeliac disease.

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Beziehung zwischen der Gliadin-Peptid-Struktur und ihr Einfluß auf den fetalen Kückendarm

Zusammenfassung Die Tendenz zur Bildung einer-Umwandlung im-Gliadin wurde bei Anwendung eines mathematischen Programms zur Vorhersage von B-Zelldeterminanten bestimmt, welches auf der Wahrscheinlichkeit der-Umwandlung nach Chou und Fasman basiert. Es wurden 6 Sequenzen gefunden, die eine hohe Wahrscheinlichkeit für die Bildung von-Umwandlungen aufwiesen. Zwischen diesen 6 Sequenzen und 3 Regionen im-Gliadin mit der Sequenz Pro-Ser-Gln-Gln, die kürzlich als verantwortlich für die Toxizität bei Cöliakie angesehen wurden, konnte eine statistisch gesicherte Beziehung gefunden werden. Mittels Festphasensynthese wurden 7 Peptide erhalten, die die oben genannten Regionen überdeckten. Ihre Toxizität wurde im fetalen Kückendarm getestet. Die Ergebnisse weisen darauf hin, daß Peptide, welche die Sequenz Pro-Ser-Gln-Gln und Gln-Gln-Gln-Pro enthalten, an der Pathogenese der Cöliakie beteiligt sein könnten.

     

Copper Transfer from Cu–Aβ to Human Serum Albumin Inhibits Aggregation,Radical Production and Reduces Aβ Toxicity

Amyloid‐β peptides (Aβ) and the protein human serum albumin (HSA) interact in vivo. They are both localised in the blood plasma and in the cerebrospinal fluid. Among other functions, HSA is involved in the transport of the essential metal copper. Complexes between Aβ and copper ions have been proposed to be an aberrant interaction implicated in the development of Alzheimer's disease, where Cu is involved in Aβ aggregation and production of reactive oxygen species (ROS). In the present work, we studied copper‐exchange reaction between Aβ and HSA or the tetrapeptide DAHK (N‐terminal Cu‐binding domain of HSA) and the consequence of this exchange on Aβ‐induced ROS production and cell toxicity. The following results were obtained: 1) HSA and DAHK removed Cu^II from Aβ rapidly and stoichiometrically, 2) HSA and DAHK were able to decrease Cu‐induced aggregation of Aβ, 3) HSA and DAHK suppressed the catalytic HO^. production in vitro and ROS production in neuroblastoma cells generated by Cu–Aβ and ascorbate, 4) HSA and DAHK were able to rescue these cells from the toxicity of Cu–Aβ with ascorbate, 5) DAHK was more potent in ROS suppression and restoration of neuroblastoma cell viability than HSA, in correlation with an easier reduction of Cu^II–HSA than Cu–DAHK by ascorbate, in vitro. Our data suggest that HSA is able to decrease aberrant Cu^II–Aβ interaction. The repercussion of the competition between HSA and Aβ to bind Cu in the blood and brain and its relation to Alzheimer's disease are discussed.     

Isolation of faba bean legumin — a comparative study of various methods

Different methods of legumin isolation from faba beans were compared with regard to the removal of vicilin taking into consideration the additional influence of variety. SDS-PAGE, analytical ultracentrifugation and SE-HPLC were used for the analysis of protein composition. SDS-PAGE analysis of the isolated legumin samples revealed, that neither combined salt fraction/isoelectric precipitation nor heating to 73 °C resulted in a complete removal of vicilin. Time-consuming isoelectric zonal precipitation and heating at 83 °C as described by Schlesier et al. (Biochem. Physiol. Pflanzen 180 (1985) 225) provides legumin preparations free of vicilin in low yields. Acceptable yields in shorter time were obtained by combining isoelectric precipitation/salt fractionation with Schlesier's procedure. SDS-PAGE analysis showed a certain dependence on variety of the patterns of legumin α-polypeptide chains.     

Intravital FRET: Probing Cellular and Tissue Function in Vivo

The development of intravital Förster Resonance Energy Transfer (FRET) is required to probe cellular and tissue function in the natural context: the living organism. Only in this way can biomedicine truly comprehend pathogenesis and develop effective therapeutic strategies. Here we demonstrate and discuss the advantages and pitfalls of two strategies to quantify FRET in vivo—ratiometrically and time-resolved by fluorescence lifetime imaging—and show their concrete application in the context of neuroinflammation in adult mice.     

Selected physico-chemical properties of succinylated legumin from pea (Pisum sativum L.)

Selected physico-chemical properties of pea legumin before and after succinylation have been investigated using isoelectric focusing, PAGE, SDS-PAGE, hydrophobicity measurements, SE-HPLC and RP-HPLC. Exhaustive succinylation shifted the I.P. of legumin from 4.75 to 3.5. The stepwise dissociation of legumin by increasing succinylation has been confirmed both by means of PAGE in a nondenaturing system, and by SE-HPLC. The results of SDS-PAGE provided evidence for the exposure of α-polypeptide chains in the native legumin. High succinylation resulted in a decrease of the surface hydrophobicity (S₀) measured by both fluorescence probes (cis-parinaric acid and anilino-naphtalene sulfonic acid). RP-HPLC gave a response both to conformational changes and the introduced succinyl residues.