Copyright © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
February 09, 2015
Very Important Paper: Establishing the Stability and Reversibility of Protein Pyrophosphorylation with Synthetic Peptides
Lisa M. Yates and Dorothea Fiedler*
Key factors in elucidating the biological relevance of novel post-translational modifications are to understand their chemical and biochemical stability as well as their reversibility in a cellular context. To evaluate these factors for protein pyrophosphorylation (a currently poorly characterized modification) Yates and Fiedler (Princeton University) used a series of synthetic pyrophosphopeptides to demonstrate that the pyrophosphate moiety is generally chemically inert but can be rapidly removed by alkaline phosphatases in vitro. Most importantly, they observed enzyme-dependent depyrophosphorylation of the synthetic pyrophosphopeptides in mammalian and yeast cell lysates. These findings thus provide the initial evidence for the reversibility of pyrophosphorylation and highlight the potential impact of this modification on cellular signaling networks.
Recently Published Articles
- Catalysing Mirror Life
Dr. Marleen Renders and Dr. Vitor B. Pinheiro
Article first published online: 4 MAR 2015 | DOI: 10.1002/cbic.201500010
Origin of life: Although current life is homochiral (with D nucleic acids), little is known about how homochirality emerged or even if it was a necessary step. The isolation of cross-chiral nucleic acid ligases demonstrate that an early heterochiral life could have been possible.
- In Vitro Reconstitution of a PKS Pathway for the Biosynthesis of Galbonolides in Streptomyces sp. LZ35
Dr. Chao Liu, Dr. Jing Zhu, Dr. Yaoyao Li, Juanli Zhang, Dr. Chunhua Lu, Dr. Haoxin Wang and Prof. Dr. Yuemao Shen
Article first published online: 3 MAR 2015 | DOI: 10.1002/cbic.201500017
Biosynthetic beauty: The gbn gene cluster encodes a hybrid FAS-PKS pathway and is involved in the biosynthesis of galbonolides, 14-membered macrolides, in Streptomyces sp. LZ35. In vitro reconstitution successfully synthesized the heptaketide skeleton of galbonolide B by using purified enzymes GbnA–C. This study provides new insights into the evolution of polyketide biosynthetic gene clusters.
- Improving Cell-Free Protein Synthesis through Genome Engineering of Escherichia coli Lacking Release Factor 1
Dr. Seok Hoon Hong, Dr. Yong-Chan Kwon, Rey W. Martin, Benjamin J. Des Soye, Alexandra M. de Paz, Kirsten N. Swonger, Dr. Ioanna Ntai, Prof. Neil L. Kelleher and Prof. Michael C. Jewett
Article first published online: 3 MAR 2015 | DOI: 10.1002/cbic.201402708
Cell-free synthetic biology: We have developed a high-yielding, cell-free protein synthesis (CFPS) platform for site-specific incorporation of non-standard amino acids (NSAAs) into proteins. Multiplex automated genome engineering (MAGE) was used to construct RF1deficient strains with improved performance, thereby opening new avenues for using cell-free synthetic biology for synthetic chemistry.
- Structures of the Apo and FAD-Bound Forms of 2-Hydroxybiphenyl 3-monooxygenase (HbpA) Locate Activity Hotspots Identified by Using Directed Evolution
Dr. Chantel N. Jensen, Tamara Mielke, Joseph E. Farrugia, Dr. Annika Frank, Henry Man, Sam Hart, Dr. Johan P. Turkenburg and Prof. Dr. Gideon Grogan
Article first published online: 3 MAR 2015 | DOI: 10.1002/cbic.201402701
Spotting hotspots: HbpA is a model for the application and suitability of flavoprotein hydroxylases, but its structure has proved elusive. We present the first apo and FAD-containing structures of HbpA, which reveal determinants of flavin binding, putative catalytic residues and also locate mutational loci previously identified by directed evolution; these could improve activity and broaden substrate range.
- Covalent and Stable CuAAC Modification of Silicon Surfaces for Control of Cell Adhesion
Surendra Vutti, Nina Buch-Månson, Dr. Sanne Schoffelen, Prof. Nicolas Bovet, Prof. Karen L. Martinez and Prof. Morten Meldal
Article first published online: 3 MAR 2015 | DOI: 10.1002/cbic.201402629
Stuck tight: The fully optimized coating of silicon surfaces with a monolayer of (3-azidopropyl)siloxide allowed attachment of secondary complex molecules. D-Amino acid adhesion peptides k-l-h-r-l-r-a and k-l-y-r-v-r-a were immobilized through CuAAC, and their interaction with cells was investigated by flow shear. Stable adhesion of HEK293 cells was achieved even at high flow rates.