Peptide specificity and lipid activation of the lysosomal transport complex ABCB9 (TAPL)

The lysosomal ABC transporter associated with antigen processing-like (TAPL, ABCB9) acts as an ATP-dependent polypeptide transporter with broad length selectivity. To characterize in detail its substrate specificity, a procedure for functional reconstitution of human TAPL was developed. By intensive screening of detergents, ideal solubilization conditions were evolved with respect to efficiency, long term stability, and functionality of TAPL. TAPL was isolated in a two-step procedure with high purity and, subsequently, reconstituted into proteoliposomes. The peptide transport activity of reconstituted TAPL strongly depends on the lipid composition. With the help of combinatorial peptide libraries, the key positions of the peptides were localized to the N- and C-terminal residues with respect to peptide transport. At both ends, TAPL favors positively charged, aromatic, or hydrophobic residues and disfavors negatively charged residues as well as asparagine and methionine. Besides specific interactions of both terminal residues, electrostatic interactions are important, since peptides with positive net charge are more efficiently transported than negatively charged ones.     

Imperceptible senescence: ageing in the ocean quahog Arctica islandica

The ocean quahog Arctica islandica is the longest-lived of all bivalve and molluscan species on earth. Animals close to 400 years are common and reported maximum live span around Iceland is close to 400 years. High and stable antioxidant capacities are a possible strategy to slow senescence and extend lifespan and this study has investigated several antioxidant parameters and a mitochondrial marker enzyme in a lifetime range spanning from 4-200 years in the Iceland quahog. In gill and mantle tissues of 4-192 year old A. islandica, catalase, citrate synthase activity and glutathione concentration declined rapidly within the first 25 years, covering the transitional phase of rapid somatic growth and sexual maturation to the outgrown mature stages (approximately 32 years). Thereafter all three parameters kept rather stable levels for > 150 years. In contrast, superoxide dismutase activities maintained high levels throughout life time. These findings support the 'Free Radical-Rate of Living theory', antioxidant capacities of A. islandica are extraordinarily high and thus may explain the species long life span.     

The TAP translocation machinery in adaptive immunity and viral escape mechanisms

The adaptive immune system plays an essential role in protecting vertebrates against a broad range of pathogens and cancer. The MHC class I-dependent pathway of antigen presentation represents a sophisticated cellular machinery to recognize and eliminate infected or malignantly transformed cells, taking advantage of the proteasomal turnover of the cell's proteome. TAP (transporter associated with antigen processing) 1/2 (ABCB2/3, where ABC is ATP-binding cassette) is the principal component in the recognition, translocation, chaperoning, editing and final loading of antigenic peptides on to MHC I complexes in the ER (endoplasmic reticulum) lumen. These different tasks are co-ordinated within a dynamic macromolecular peptide-loading complex consisting of TAP1/2 and various auxiliary factors, such as the adapter protein tapasin, the oxidoreductase ERp57, the lectin chaperone calreticulin, and the final peptide acceptor the MHC I heavy chain associated with β2-microglobulin. In this chapter, we summarize the structural organization and molecular mechanism of the antigen-translocation machinery as well as various modes of regulation by viral factors and in genetic diseases and tumour development.     

Zur Kenntnis der Zell- und Kernteilung in dem primären Meristem

Ohne Zusammenfassung     

Cysteine sulfinic acid uptake in cultured neuronal and glial cells

The presence of an efficient high affinity uptake system for L-CSA has been demonstrated in cultured neuronal and glial cells of various types. In neurons and most glial cells L-CSA uptake is inhibited by acidic amino acids,L-glutamate andL-aspartate and requires sodium ions; however the sodium dependence varies from one cell type to the other. The characteristics of the uptake system change during cell maturation, especially in astroblasts. The predominance of CSA uptake in glial cells as compared to neurons, the similarity of the kinetic parameters and of the structural specficity ofL-glutamate uptake suggest that both excitatory amino acids are transported by a common system. In view of accumulating evidence, the present results are in agreement with a role of CSA as a neurotransmitter and as a precursor for taurine biosynthesis in the central nervous system.     

Carotenoids in petals of perennial Medicago species

The carotenoid content in the petals of fourteen Medicago species was examined, together with eight species studied previously, caratenoids in all the known perennials of the genus are reported. The species can be arranged in relationship groups on the basis of their interfertility. No major carotenoid was species- or groupspecific; a few minor pigments, however, were group- or species-specific. The amount of carotenoids ranged from 7 μg/g dry matter in violet-flowered M. sativa to 2120 μg/g in brownish-yellow M. platycarpos. Xanthophylls constituted 76–99% of the total, with lutein as the major component. β-Carotene, lutein and flavoxanthin were ubiquitous in petals. In M sativa leaves β-carotene, lutein, violaxanthin and neoxanthin constituted 88% of the total. The xanthophylls were esterified in petals but not in leaves.     

Structure and dynamics of membrane-associated ICP47, a viral inhibitor of the MHC I antigen-processing machinery

To evade the host's immune response, herpes simplex virus employs the immediate early gene product ICP47 (IE12) to suppress antigen presentation to cytotoxic T-lymphocytes by inhibition of the ATP-binding cassette transporter associated with antigen processing (TAP). ICP47 is a membrane-associated protein adopting an alpha-helical conformation. Its active domain was mapped to residues 3-34 and shown to encode all functional properties of the full-length protein. The active domain of ICP47 was reconstituted into oriented phospholipid bilayers and studied by proton-decoupled 15N and 2H solid-state NMR spectroscopy. In phospholipid bilayers, the protein adopts a helix-loop-helix structure, where the average tilt angle of the helices relative to the membrane surface is approximately 15 degrees (+/- 7 degrees ). The alignment of both structured domains exhibits a mosaic spread of approximately 10 degrees . A flexible dynamic loop encompassing residues 17 and 18 separates the two helices. Refinement of the experimental data indicates that helix 1 inserts more deeply into the membrane. These novel insights into the structure of ICP47 represent an important step toward a molecular understanding of the immune evasion mechanism of herpes simplex virus and are instrumental for the design of new therapeutics.