Journal Issue - Volume 17 Issue 4 (April 2008)
Induced‐fit or preexisting equilibrium dynamics? Lessons from protein crystallography and MD simulations on acetylcholinesterase and implications for structure‐based drug design
- Yechun Xu, Jacques Ph. Colletier, Hualiang Jiang, Israel Silman, Joel L. Sussman, Martin Weik
- Published in Wiley Interscience on Jan 02, 2009
- DOI: 10.1110/ps.083453808 (p 601-605)
Abstract Crystal structures of acetylcholinesterase complexed with ligands are compared with side‐chain conformations accessed by native acetylcholinesterase in molecular dynamics (MD) simulations. Several crystallographic conformations of a key residue in a specific binding site are accessed in a simulation of native acetylcholinesterase, although not seen in rotomer plots. Conformational changes upon ligand binding thus involve...
The epitope space of the human proteome
- Lisa Berglund, Jorge Andrade, Jacob Odeberg, Mathias Uhlén
- Published in Wiley Interscience on Jan 02, 2009
- DOI: 10.1110/ps.073347208 (p 606-613)
Abstract In the post‐genome era, there is a great need for protein‐specific affinity reagents to explore the human proteome. Antibodies are suitable as reagents, but generation of antibodies with low cross‐reactivity to other human proteins requires careful selection of antigens. Here we show the results from a proteome‐wide effort to map linear epitopes based on uniqueness relative to the entire human proteome. The analysis was...
Design of a highly specific and noninvasive biosensor suitable for real‐time in vivo imaging of mercury (II) uptake
- Richard R. Chapleau, Rebecca Blomberg, Peter C. Ford, Martin Sagermann
- Published in Wiley Interscience on Jan 02, 2009
- DOI: 10.1110/ps.073358908 (p 614-622)
Abstract Mercury is a ubiquitous pollutant that when absorbed is extremely toxic to a wide variety of biochemical processes. Mercury (II) is a strong, “invisible” poison that is rapidly absorbed by tissues of the intestinal tract, kidneys, and liver upon ingestion. In this study, a novel fluorescence‐based biosensor is presented that allows for the direct monitoring of the uptake and distribution of the metal under noninvasive in...
Analysis of solvent content and oligomeric states in protein crystals—does symmetry matter?
- Maksymilian Chruszcz, Wojciech Potrzebowski, Matthew D. Zimmerman, Marek Grabowski, Heping Zheng, Piotr Lasota, Wladek Minor
- Published in Wiley Interscience on Jan 02, 2009
- DOI: 10.1110/ps.073360508 (p 623-632)
Abstract A nonredundant set of 9081 protein crystal structures in the Protein Data Bank was used to examine the solvent content, the number of polypeptide chains, and the oligomeric states of proteins in crystals as a function of crystal symmetry (as classified by crystal systems and space groups). It was found that there is a correlation between solvent content and crystal symmetry. Surprisingly, proteins crystallizing in lower...
Design of an engineered N‐terminal HIV‐1 gp41 trimer with enhanced stability and potency
- John J. Dwyer, Karen L. Wilson, Kimberly Martin, Jennifer E. Seedorff, Aisha Hasan, Robyn J. Medinas, Donna K. Davison, Michael D. Feese, Hans‐Thomas Richter, Hidong Kim, Thomas J. Matthews, Mary K. Delmedico
- Published in Wiley Interscience on Jan 02, 2009
- DOI: 10.1110/ps.073307608 (p 633-643)
Abstract HIV fusion is mediated by a conformational transition in which the C‐terminal region (HR2) of gp41 interacts with the N‐terminal region (HR1) to form a six‐helix bundle. Peptides derived from the HR1 form a well‐characterized, trimeric coiled‐coil bundle in the presence of HR2 peptides, but there is little structural information on the isolated HR1 trimer. Using protein design, we have designed synthetic HR1 peptides that...
Side chain burial and hydrophobic core packing in protein folding transition states
- Patrick J. Farber, Anthony Mittermaier
- Published in Wiley Interscience on Jan 02, 2009
- DOI: 10.1110/ps.073105408 (p 644-651)
Abstract A critical step in the folding pathway of globular proteins is the formation of a tightly packed hydrophobic core. Several mutational studies have addressed the question of whether tight packing interactions are present during the rate‐limiting step of folding. In some of these investigations, substituted side chains have been assumed to form native‐like interactions in the transition state when the folding rates of mutant...
Structural and thermodynamic effects of ANS binding to human interleukin‐1 receptor antagonist
- Ramil F. Latypov, Dingjiang Liu, Kannan Gunasekaran, Timothy S. Harvey, Vladimir I. Razinkov, Andrei A. Raibekas
- Published in Wiley Interscience on Jan 02, 2009
- DOI: 10.1110/ps.073332408 (p 652-663)
Abstract Although 8‐anilinonaphthalene‐1‐sulfonic acid (ANS) is frequently used in protein folding studies, the structural and thermodynamic effects of its binding to proteins are not well understood. Using high‐resolution two‐dimensional NMR and human interleukin‐1 receptor antagonist (IL‐1ra) as a model protein, we obtained detailed information on ANS–protein interactions in the absence and presence of urea. The effects of ambient...
Substrate specificity of human kallikreins 1 and 6 determined by phage display
- Hai‐Xin Li, Bum‐Yeol Hwang, Gurunathan Laxmikanthan, Sachiko I. Blaber, Michael Blaber, Pavel A. Golubkov, Pengyu Ren, Brent L. Iverson, George Georgiou
- Published in Wiley Interscience on Jan 02, 2009
- DOI: 10.1110/ps.073333208 (p 664-672)
Abstract The human tissue kallikrein (KLK) family contains 15 secreted serine proteases that are expressed in a wide range of tissues and have been implicated in different physiological functions and disease states. Of these, KLK1 has been shown to be involved in the regulation of multiple physiological processes such as blood pressure, smooth muscle contraction, and vascular cell growth. KLK6 is overexpressed in breast and ovarian...
Engineering membrane protein overproduction in Escherichia coli
- Daniel Martinez Molina, Tobias Cornvik, Said Eshaghi, Jesper Z. Haeggström, Pär Nordlund, Marina Ignatushchenko Sabet
- Published in Wiley Interscience on Jan 02, 2009
- DOI: 10.1110/ps.073242508 (p 673-680)
Abstract Membrane proteins play a fundamental role in human disease and therapy, but suffer from a lack of structural and functional information compared to their soluble counterparts. The paucity of membrane protein structures is primarily due to the unparalleled difficulties in obtaining detergent‐solubilized membrane proteins at sufficient levels and quality. We have developed an in vitro evolution strategy for optimizing the...
Nonspecific base recognition mediated by water bridges and hydrophobic stacking in ribonuclease I from Escherichia coli
- Sergio Martinez Rodriguez, Santosh Panjikar, Karolien Van Belle, Lode Wyns, Joris Messens, Remy Loris
- Published in Wiley Interscience on Jan 02, 2009
- DOI: 10.1110/ps.073420708 (p 681-690)
Abstract The crystal structure of Escherichia coli ribonuclease I (EcRNase I) reveals an RNase T2‐type fold consisting of a conserved core of six β‐strands and three α‐helices. The overall architecture of the catalytic residues is very similar to the plant and fungal RNase T2 family members, but the perimeter surrounding the active site is characterized by structural elements specific for E. coli. In the structure of EcRNase I in complex with a...
Crystal structure of enoyl–acyl carrier protein reductase (FabK) from Streptococcus pneumoniae reveals the binding mode of an inhibitor
- Jun Saito, Mototsugu Yamada, Takashi Watanabe, Maiko Iida, Hideo Kitagawa, Sho Takahata, Tomohiro Ozawa, Yasuo Takeuchi, Fukuichi Ohsawa
- Published in Wiley Interscience on Jan 02, 2009
- DOI: 10.1110/ps.073288808 (p 691-699)
Abstract Enoyl–acyl carrier protein (ACP) reductases are critical for bacterial type II fatty acid biosynthesis and thus are attractive targets for developing novel antibiotics. We determined the crystal structure of enoyl–ACP reductase (FabK) from Streptococcus pneumoniae at 1.7 Å resolution. There was one dimer per asymmetric unit. Each subunit formed a triose phosphate isomerase (TIM) barrel structure, and flavin mononucleotide (FMN) was...
The crystal structure of the C45S mutant of annelid Arenicola marina peroxiredoxin 6 supports its assignment to the mechanistically typical 2‐Cys subfamily without any formation of toroid‐shaped decamers
- Aude Smeets, Eléonore Loumaye, André Clippe, Jean‐François Rees, Bernard Knoops, Jean‐Paul Declercq
- Published in Wiley Interscience on Jan 02, 2009
- DOI: 10.1110/ps.073399308 (p 700-710)
Abstract The peroxiredoxins (PRDXs) define a superfamily of thiol‐dependent peroxidases able to reduce hydrogen peroxide, alkyl hydroperoxides, and peroxynitrite. Besides their cytoprotective antioxidant function, PRDXs have been implicated in redox signaling and chaperone activity, the latter depending on the formation of decameric high‐molecular‐weight structures. PRDXs have been mechanistically divided into three major...
Structure dissection of human progranulin identifies well‐folded granulin/epithelin modules with unique functional activities
- Dmitri Tolkatchev, Suneil Malik, Anna Vinogradova, Ping Wang, Zhigang Chen, Ping Xu, Hugh P.J. Bennett, Andrew Bateman, Feng Ni
- Published in Wiley Interscience on Jan 02, 2009
- DOI: 10.1110/ps.073295308 (p 711-724)
Abstract Progranulin is a secreted protein with important functions in several physiological and pathological processes, such as embryonic development, host defense, and wound repair. Autosomal dominant mutations in the progranulin gene cause frontotemporal dementia, while overexpression of progranulin promotes the invasive progression of a range of tumors, including those of the breast and the brain. Structurally, progranulin...
Structure–function studies on the active site of the coelenterazine‐dependent luciferase from Renilla
- Jongchan Woo, Matthew H. Howell, Albrecht G. von Arnim
- Published in Wiley Interscience on Jan 02, 2009
- DOI: 10.1110/ps.073355508 (p 725-735)
Abstract Renilla luciferase (RLUC) is a versatile tool for gene expression assays and in vivo biosensor applications, but its catalytic mechanism remains to be elucidated. RLUC is evolutionarily related to the α/β hydrolase family. Its closest known homologs are bacterial dehalogenases, raising the question of how a protein with a hydrolase fold can function as a decarboxylating oxygenase. Molecular docking simulations with the...
An efficient on‐column expressed protein ligation strategy: Application to segmental triple labeling of human apolipoprotein E3
- Wentao Zhao, Yonghong Zhang, Chunxian Cui, Qianqian Li, Jianjun Wang
- Published in Wiley Interscience on Jan 02, 2009
- DOI: 10.1110/ps.073383708 (p 736-747)
Abstract Expressed protein ligation (EPL) is an intein‐based approach that has been used for protein engineering and biophysical studies of protein structures. One major problem of the EPL is the low yield of final ligation product, primarily due to the complex procedure of the EPL, preventing EPL from gaining popularity in the research community. Here we report an efficient on‐column EPL strategy, which focuses on enhancing the...




