Journal Issue - Volume 18 Issue 8 (August 2009)
In this issue
- Published in Wiley Interscience on Jul 22, 2009
- DOI: 10.1002/pro.210 (p n/a-n/a)
Effect of protein structure on deamidation rate in the Fc fragment of an IgG1 monoclonal antibody
- Sandipan Sinha, Lei Zhang, Shaofeng Duan, Todd D. Williams, Josef Vlasak, Roxana Ionescu, Elizabeth M. Topp
- Published in Wiley Interscience on May 28, 2009
- DOI: 10.1002/pro.173 (p 1573-1584)
Abstract The effects of secondary structure on asparagine (N) deamidation in a 22 amino acid sequence (369‐GFYPSDIAVEWESNGQPENNYK‐390) of the crystallizable (Fc) fragment of a human monoclonal antibody (Fc IgG1) were investigated using high‐resolution ultra performance liquid chromatography with tandem mass spectrometry (UPLC/MS). Samples containing either the intact Fc IgG (∼50 kD) (“intact protein”), or corresponding synthetic...
Vibrational spectroscopic investigation of the ligand binding domain of kainate receptors
- Mei Du, Anu Rambhadran, Vasanthi Jayaraman
- Published in Wiley Interscience on May 28, 2009
- DOI: 10.1002/pro.174 (p 1585-1591)
Abstract Fourier transform infrared spectroscopy has been used to probe the agonist‐protein interactions in the ligand binding domain of the GluR6 subunit, one subunit of the kainate subtype of glutamate receptors. In order to study the changes in the interactions over a range of activations the investigations were performed using the wild type, N690S, and T661E mutations. These studies show that the strength of the interactions at...
NMR‐based characterization of a refolding intermediate of β2‐microglobulin labeled using a wheat germ cell‐free system
- Atsushi Kameda, Eugene‐Hayato Morita, Kazumasa Sakurai, Hironobu Naiki, Yuji Goto
- Published in Wiley Interscience on May 28, 2009
- DOI: 10.1002/pro.179 (p 1592-1601)
Abstract In patients with dialysis‐related amyloidosis, β2‐microglobulin (β2‐m) is a major structural component of amyloid fibrils. It has been suggested that the partial unfolding of β2‐m is a prerequisite to the formation of amyloid fibrils, and that the folding intermediate trapped by the non‐native trans‐Pro32 isomer leads to the formation of amyloid fibrils. Although clarifying the structure of this refolding intermediate by...
Alpha helical crossovers favor right‐handed supersecondary structures by kinetic trapping: The phone cord effect in protein folding
- Benjamin J. Cole, Christopher Bystroff
- Published in Wiley Interscience on Jun 08, 2009
- DOI: 10.1002/pro.182 (p 1602-1608)
Abstract The remarkable predominance of right‐handedness in beta‐alpha‐beta helical crossovers has been previously explained in terms of thermodynamic stability and kinetic accessibility, but a different kinetic trapping mechanism may also play a role. If the beta‐sheet contacts are made before the crossover helix is fully formed, and if the backbone angles of the folding helix follows the energetic pathway of least resistance, then...
High‐energy water sites determine peptide binding affinity and specificity of PDZ domains
- Thijs Beuming, Ramy Farid, Woody Sherman
- Published in Wiley Interscience on May 28, 2009
- DOI: 10.1002/pro.177 (p 1609-1619)
Abstract PDZ domains have well known binding preferences for distinct C‐terminal peptide motifs. For most PDZ domains, these motifs are of the form [S/T]‐W‐[I/L/V]. Although the preference for S/T has been explained by a specific hydrogen bond interaction with a histidine in the PDZ domain and the (I/L/V) is buried in a hydrophobic pocket, the mechanism for Trp specificity at the second to last position has thus far remained...
Detection of early unfolding events in a dimeric protein by amide proton exchange and native electrospray mass spectrometry
- James A. Mobley, Anton Poliakov
- Published in Wiley Interscience on May 28, 2009
- DOI: 10.1002/pro.176 (p 1620-1627)
Abstract Oligomeric proteins generally undergo unfolding through a dissociation/denaturation mechanism wherein the subunits first dissociate and then unfold. This mechanism can be detected by the fact that the proteins exhibit a concentration dependence of the denaturation curve. However, the concentration dependence does not answer the question of whether there are thermally induced conformational changes that facilitate subunit...
Variants of human thymidylate synthase with loop 181–197 stabilized in the inactive conformation
- Leslie L. Lovelace, Saphronia R. Johnson, Lydia M. Gibson, Brittnaie J. Bell, Sondra H. Berger, Lukasz Lebioda
- Published in Wiley Interscience on May 28, 2009
- DOI: 10.1002/pro.171 (p 1628-1636)
Abstract Loop 181–197 of human thymidylate synthase (hTS) populates two major conformations, essentially corresponding to the loop flipped by 180°. In one of the conformations, the catalytic Cys195 residue lies distant from the active site making the enzyme inactive. Ligands stabilizing this inactive conformation may function as allosteric inhibitors. To facilitate the search for such inhibitors, we have expressed and characterized...
Osmolyte perturbation reveals conformational equilibria in spin‐labeled proteins
- Carlos J. López, Mark R. Fleissner, Zhefeng Guo, Ana K. Kusnetzow, Wayne L. Hubbell
- Published in Wiley Interscience on Jun 08, 2009
- DOI: 10.1002/pro.180 (p 1637-1652)
Abstract Recent evidence suggests that proteins at equilibrium can exist in a manifold of conformational substates, and that these substates play important roles in protein function. Therefore, there is great interest in identifying regions in proteins that are in conformational exchange. Electron paramagnetic resonance spectra of spin‐labeled proteins containing the nitroxide side chain (R1) often consist of two (or more)...
Structure and stability of icosahedral particles of a covalent coat protein dimer of bacteriophage MS2
- Pavel Plevka, Kaspars Tars, Lars Liljas
- Published in Wiley Interscience on Jun 11, 2009
- DOI: 10.1002/pro.184 (p 1653-1661)
Abstract Particles formed by the bacteriophage MS2 coat protein mutants with insertions in their surface loops induce a strong immune response against the inserted epitopes. The covalent dimers created by fusion of two copies of the coat protein gene are more tolerant to various insertions into the surface loops than the single subunits. We determined a 4.7‐Å resolution crystal structure of an icosahedral particle assembled from...
Two alternative modes for optimizing nylon‐6 byproduct hydrolytic activity from a carboxylesterase with a β‐lactamase fold: X‐ray crystallographic analysis of directly evolved 6‐aminohexanoate‐dimer hydrolase
- Taku Ohki, Naoki Shibata, Yoshiki Higuchi, Yasuyuki Kawashima, Masahiro Takeo, Dai‐ichiro Kato, Seiji Negoro
- Published in Wiley Interscience on Jun 11, 2009
- DOI: 10.1002/pro.185 (p 1662-1673)
Abstract Promiscuous 6‐aminohexanoate‐linear dimer (Ald)‐hydrolytic activity originally obtained in a carboxylesterase with a β‐lactamase fold was enhanced about 80‐fold by directed evolution using error‐prone PCR and DNA shuffling. Kinetic studies of the mutant enzyme (Hyb‐S4M94) demonstrated that the enzyme had acquired an increased affinity (Km = 15 mM) and turnover (kcat = 3.1 s−1) for Ald, and that a catalytic center suitable for nylon‐6...
Exploring functional roles of multibinding protein interfaces
- Manoj Tyagi, Benjamin A. Shoemaker, Stephen H. Bryant, Anna R. Panchenko
- Published in Wiley Interscience on Jun 08, 2009
- DOI: 10.1002/pro.181 (p 1674-1683)
Abstract Cellular processes are highly interconnected and many proteins are shared in different pathways. Some of these shared proteins or protein families may interact with diverse partners using the same interface regions; such multibinding proteins are the subject of our study. The main goal of our study is to attempt to decipher the mechanisms of specific molecular recognition of multiple diverse partners by promiscuous protein...
Homology modeling of human Toll‐like receptors TLR7, 8, and 9 ligand‐binding domains
- Tiandi Wei, Jing Gong, Ferdinand Jamitzky, Wolfgang M. Heckl, Robert W. Stark, Shaila C. Rössle
- Published in Wiley Interscience on Jun 11, 2009
- DOI: 10.1002/pro.186 (p 1684-1691)
Abstract Toll‐like receptors (TLRs) play a key role in the innate immune system. The TLR7, 8, and 9 compose a family of intracellularly localized TLRs that signal in response to pathogen‐derived nucleic acids. So far, there are no crystallographic structures for TLR7, 8, and 9. For this reason, their ligand‐binding mechanisms are poorly understood. To enable first predictions of the receptor–ligand interaction sites, we developed...
Native like structure in the unfolded state of the villin headpiece helical subdomain, an ultrafast folding protein
- Wenli Meng, Bing Shan, Yuefeng Tang, Daniel P. Raleigh
- Published in Wiley Interscience on May 06, 2009
- DOI: 10.1002/pro.152 (p 1692-1701)
Abstract The villin headpiece subdomain, HP36, is the smallest naturally occurring protein that folds cooperatively. Its small size, rapid folding, and simple three‐helix topology have made it an extremely popular system for computational studies of protein folding. The role of unfolded state structure in rapid folding is an area of active investigation, but relatively little is known about the properties of unfolded states under...
Molecular modeling of flexible arm‐mediated interactions between bacterial chemoreceptors and their modification enzyme
- Usha K. Muppirala, Susan Desensi, Terry P. Lybrand, Gerald L. Hazelbauer, Zhijun Li
- Published in Wiley Interscience on May 28, 2009
- DOI: 10.1002/pro.170 (p 1702-1714)
Abstract Sensory adaptation in bacterial chemotaxis is mediated by methylation and demethylation of specific glutamyl residues in the cytoplasmic domain of chemoreceptors. Methylation is catalyzed by methyltransferase CheR. In E. coli and related organisms, methylation sufficiently rapid to be physiologically effective requires a carboxyl terminal pentapeptide sequence on the receptor being modified or, via adaptational assistance, on a...




