@Article{IPB-816, author = {Nishiyama, T. and Sakayama, H. and de Vries, J. and Buschmann, H. and Saint-Marcoux, D. and Ullrich, K. K. and Haas, F. B. and Vanderstraeten, L. and Becker, D. and Lang, D. and Vosolsobě, S. and Rombauts, S. and Wilhelmsson, P. K. and Janitza, P. and Kern, R. and Heyl, A. and Rümpler, F. and Calderón Villalobos, L. I. A. and Clay, J. M. and Skokan, R. and Toyoda, A. and Suzuki, Y. and Kagoshima, H. and Schijlen, E. and Tajeshwar, N. and Catarino, B. and Hetherington, A. J. and Saltykova, A. and Bonnot, C. and Breuninger, H. and Symeonidi, A. and Radhakrishnan, G. V. and Van Nieuwerburgh, F. and Deforce, D. and Chang, C. and Karol, K. G. and Hedrich, R. and Ulvskov, P. and Glöckner, G. and Delwiche, C. F. and Petrášek, J. and Van de Peer, Y. and Friml, J. and Beilby, M. and Dolan, L. and Kohara, Y. and Sugano, S. and Fujiyama, A. and Delaux, P.-M. and Quint, M. and Theißen, G. and Hagemann, M. and Harholt, J. and Dunand, C. and Zachgo, S. and Langdale, J. and Maumus, F. and Van Der Straeten, D. and Gould, S. B. and Rensing, S. A. and}, title = {{The Chara Genome: Secondary Complexity and Implications for Plant Terrestrialization}}, year = {2018}, pages = {448-464.e24}, journal = {Cell}, doi = {10.1016/j.cell.2018.06.033}, volume = {174}, abstract = {Land plants evolved from charophytic algae, among which Charophyceae possess the most complex body plans. We present the genome of Chara braunii; comparison of the genome to those of land plants identified evolutionary novelties for plant terrestrialization and land plant heritage genes. C. braunii employs unique xylan synthases for cell wall biosynthesis, a phragmoplast (cell separation) mechanism similar to that of land plants, and many phytohormones. C. braunii plastids are controlled via land-plant-like retrograde signaling, and transcriptional regulation is more elaborate than in other algae. The morphological complexity of this organism may result from expanded gene families, with three cases of particular note: genes effecting tolerance to reactive oxygen species (ROS), LysM receptor-like kinases, and transcription factors (TFs). Transcriptomic analysis of sexual reproductive structures reveals intricate control by TFs, activity of the ROS gene network, and the ancestral use of plant-like storage and stress protection proteins in the zygote.} } @Article{IPB-2184, author = {Schilling, S. and Hoffmann, T. and Rosche, F. and Manhart, S. and Wasternack, C. and Demuth, H.-U. and}, title = {{Heterologous Expression and Characterization of Human Glutaminyl Cyclase: Evidence for a Disulfide Bond with Importance for Catalytic Activity}}, year = {2002}, pages = {10849-10857}, journal = {Biochemistry}, doi = {10.1021/bi0260381}, volume = {41}, abstract = {Glutaminyl cyclase (QC, EC 2.3.2.5) catalyzes the formation of pyroglutamate residues from glutamine at the N-terminus of peptides and proteins. In the current study, human QC was functionally expressed in the secretory pathway of Pichia pastoris, yielding milligram quantities after purification from the supernatant of a 5 L fermentation. Initial characterization studies of the recombinant QC using MALDI-TOF mass spectrometry revealed correct proteolytic processing and N-glycosylation at both potential sites with similar 2 kDa extensions. CD spectral analysis indicated a high α-helical content, which contrasts with plant QC from Carica papaya. The kinetic parameters for conversion of H-Gln-Tyr-Ala-OH by recombinant human QC were almost identical to those previously reported for purified bovine pituitary QC. However, the results obtained for conversion of H-Gln-Gln-OH, H-Gln-NH2, and H-Gln-AMC were found to be contradictory to previous studies on human QC expressed intracellularly in E. coli. Expression of QC in E. coli showed that approximately 50% of the protein did not contain a disulfide bond that is present in the entire QC expressed in P. pastoris. Further, the enzyme was consistently inactivated by treatment with 15 mM DTT, whereas deglycosylation had no effect on enzymatic activity. Analysis of the fluorescence spectra of the native, reduced, and unfolded human QC point to a conformational change of the protein upon treatment with DTT. In terms of the different enzymatic properties, the consequences of QC expression in different environments are discussed.} }