Journal Issue - Volume 19 Issue 8 (August 2010)
In This Issue
- Published in Wiley Interscience on May 27, 2010
- DOI: 10.1002/pro.452 (p )
A cytochrome c fusion protein domain for convenient detection, quantification, and enhanced production of membrane proteins in Escherichia coli —Expression and characterization of cytochrome‐tagged Complex I subunits
- Tobias Gustavsson, Maria Trane, Vamsi K. Moparthi, Egle Miklovyte, Lavanya Moparthi, Kamil Górecki, Thom Leiding, Sindra Peterson Årsköld, Cecilia Hägerhäll
- Published in Wiley Interscience on Jun 29, 2010
- DOI: 10.1002/pro.424 (p 1445-1460)
Abstract Overproduction of membrane proteins can be a cumbersome task, particularly if high yields are desirable. NADH:quinone oxidoreductase (Complex I) contains several very large membrane‐spanning protein subunits that hitherto have been impossible to express individually in any appreciable amounts in Escherichia coli. The polypeptides contain no prosthetic groups and are poorly antigenic, making optimization of protein production a...
Domain distribution and intrinsic disorder in hubs in the human protein–protein interaction network
- Ashwini Patil, Kengo Kinoshita, Haruki Nakamura
- Published in Wiley Interscience on Jun 29, 2010
- DOI: 10.1002/pro.425 (p 1461-1468)
Abstract Intrinsic disorder and distributed surface charge have been previously identified as some of the characteristics that differentiate hubs (proteins with a large number of interactions) from non‐hubs in protein–protein interaction networks. In this study, we investigated the differences in the quantity, diversity, and functional nature of Pfam domains, and their relationship with intrinsic disorder, in hubs and non‐hubs. We...
High resolution X‐ray structures of mouse major urinary protein nasal isoform in complex with pheromones
- Samantha Perez‐Miller, Qin Zou, Milos V. Novotny, Thomas D. Hurley
- Published in Wiley Interscience on Jun 29, 2010
- DOI: 10.1002/pro.426 (p 1469-1479)
Abstract In mice, the major urinary proteins (MUP) play a key role in pheromonal communication by binding and transporting semiochemicals. MUP‐IV is the only isoform known to be expressed in the vomeronasal mucosa. In comparison with the MUP isoforms that are abundantly excreted in the urine, MUP‐IV is highly specific for the male mouse pheromone 2‐sec‐butyl‐4,5‐dihydrothiazole (SBT). To examine the structural basis of this ligand...
X‐ray crystallographic analysis of adipocyte fatty acid binding protein (aP2) modified with 4‐hydroxy‐2‐nonenal
- Kristina Hellberg, Paul A. Grimsrud, Andrew C. Kruse, Leonard J. Banaszak, Douglas H. Ohlendorf, David A. Bernlohr
- Published in Wiley Interscience on Jun 29, 2010
- DOI: 10.1002/pro.427 (p 1480-1489)
Abstract Fatty acid binding proteins (FABP) have been characterized as facilitating the intracellular solubilization and transport of long‐chain fatty acyl carboxylates via noncovalent interactions. More recent work has shown that the adipocyte FABP is also covalently modified in vivo on Cys117 with 4‐hydroxy‐2‐nonenal (4‐HNE), a bioactive aldehyde linked to oxidative stress and inflammation. To evaluate 4‐HNE binding and modification, the...
Circularly permuted monomeric red fluorescent proteins with new termini in the β‐sheet
- Haley J. Carlson, Darrel W. Cotton, Robert E. Campbell
- Published in Wiley Interscience on Jul 08, 2010
- DOI: 10.1002/pro.428 (p 1490-1499)
Abstract Circularly permuted fluorescent proteins (FPs) have a growing number of uses in live cell fluorescence biosensing applications. Most notably, they enable the construction of single fluorescent protein‐based biosensors for Ca2+ and other analytes of interest. Circularly permuted FPs are also of great utility in the optimization of fluorescence resonance energy transfer (FRET)‐based biosensors by providing a means for varying the...
Effect of ligand binding on human
D ‐amino acid oxidase: Implications for the development of new drugs for schizophrenia treatment- Laura Caldinelli, Gianluca Molla, Luisa Bracci, Barbara Lelli, Silvia Pileri, Pamela Cappelletti, Silvia Sacchi, Loredano Pollegioni
- Published in Wiley Interscience on Jul 08, 2010
- DOI: 10.1002/pro.429 (p 1500-1512)
Abstract In human brain the flavoprotein D‐amino acid oxidase (hDAAO) is responsible for the degradation of the neuromodulator D‐serine, an important effector of NMDA‐receptor mediated neurotransmission. Experimental evidence supports the concept that D‐serine concentration increase by hDAAO inhibition may represent a valuable therapeutic approach to improve the symptoms in schizophrenia patients. This study investigated the effects on hDAAO...
Crystal structures of the apo and ATP bound Mycobacterium tuberculosis nitrogen regulatory PII protein
- Nishant D. Shetty, Manchi C. M. Reddy, Satheesh K. Palaninathan, Joshua L. Owen, James C. Sacchettini
- Published in Wiley Interscience on Jul 09, 2010
- DOI: 10.1002/pro.430 (p 1513-1524)
Abstract PII constitutes a family of signal transduction proteins that act as nitrogen sensors in microorganisms and plants. Mycobacterium tuberculosis (Mtb) has a single homologue of PII whose precise role has as yet not been explored. We have solved the crystal structures of the Mtb PII protein in its apo and ATP bound forms to 1.4 and 2.4 Å resolutions, respectively. The protein forms a trimeric assembly in the crystal lattice and folds...
The Deinococcus radiodurans Snf2 intein caught in the act: Detection of the Class 3 intein signature Block F branched intermediate
- Lear E. Brace, Maurice W. Southworth, Kazuo Tori, Michelle L. Cushing, Francine Perler
- Published in Wiley Interscience on Jun 29, 2010
- DOI: 10.1002/pro.431 (p 1525-1533)
Abstract Inteins are the protein equivalent of introns. They are remarkable and robust single turnover enzymes that splice out of precursor proteins during post‐translational maturation of the host protein (extein). The Deinococcus radiodurans Snf2 intein is the second member of the recently discovered Class 3 subfamily of inteins to be characterized. Class 3 inteins have a unique sequence signature: (a) they start with residues other than the...
Use of protein trans‐splicing to produce active and segmentally 2 H, 15 N labeled mannuronan C5‐epimerase AlgE4
- Edith Buchinger, Finn L. Aachmann, A. Sesilja Aranko, Svein Valla, Gudmund Skjåk‐BræK, Hideo Iwaï, Reinhard Wimmer
- Published in Wiley Interscience on May 27, 2010
- DOI: 10.1002/pro.432 (p 1534-1543)
Abstract Alginate epimerases are large multidomain proteins capable of epimerising C5 on β‐D‐mannuronic acid (M) turning it into α‐L‐guluronic acid (G) in a polymeric alginate. Azotobactervinelandii secretes a family of seven epimerases, each of which is capable of producing alginates with characteristic G distribution patterns. All seven epimerases consist of two types of modules, denoted A and R, in varying numbers. Attempts to study these ...
Protein denaturation and protein:drugs interactions from intrinsic protein fluorescence measurements at the nanolitre scale
- Gaudet Matthieu, Remtulla Nina, Jackson Sophie E., Main Ewan R. G., Bracewell Daniel G., Aeppli Gabriel, Dalby Paul A.
- Published in Wiley Interscience on Jul 13, 2010
- DOI: 10.1002/pro.433 (p 1544-1554)
Abstract Protein stability and ligand‐binding affinity measurements are widely required for the formulation of biopharmaceutical proteins, protein engineering and drug screening within life science research. Current techniques either consume too much of often precious biological or compound materials, in large sample volumes, or alternatively require chemical labeling with fluorescent tags to achieve measurements at submicrolitre...
Temperature‐dependent structural changes in intrinsically disordered proteins: Formation of α‒helices or loss of polyproline II?
- Magnus Kjaergaard, Ann‒Beth Nørholm, Ruth Hendus‒Altenburger, Stine F. Pedersen, Flemming M. Poulsen, Birthe B. Kragelund
- Published in Wiley Interscience on Jul 13, 2010
- DOI: 10.1002/pro.435 (p 1555-1564)
Abstract Structural characterization of intrinsically disordered proteins (IDPs) is mandatory for deciphering their potential unique physical and biological properties. A large number of circular dichroism (CD) studies have demonstrated that a structural change takes place in IDPs with increasing temperature, which most likely reflects formation of transient α‒helices or loss of polyproline II (PPII) content. Using three IDPs, ACTR,...
Crystal structure and kinetic mechanism of aminoglycoside phosphotransferase‐2″‐IVa
- Marta Toth, Hilary Frase, Nuno Tiago Antunes, Clyde A. Smith, Sergei B. Vakulenko
- Published in Wiley Interscience on Jul 13, 2010
- DOI: 10.1002/pro.437 (p 1565-1576)
Abstract Acquired resistance to aminoglycoside antibiotics primarily results from deactivation by three families of aminoglycoside‐modifying enzymes. Here, we report the kinetic mechanism and structure of the aminoglycoside phosphotransferase 2″‐IVa (APH(2″)‐IVa), an enzyme responsible for resistance to aminoglycoside antibiotics in clinical enterococcal and staphylococcal isolates. The enzyme operates via a Bi‐Bi sequential...
NMR and crystallographic structures of the FK506 binding domain of human malarial parasite Plasmodium vivax FKBP35
- Reema Alag, Insaf A. Qureshi, Nagakumar Bharatham, Joon Shin, Julien Lescar, Ho Sup Yoon
- Published in Wiley Interscience on May 27, 2010
- DOI: 10.1002/pro.438 (p 1577-1586)
Abstract The emergence of drug‐resistant malaria parasites is the major threat to effective malaria control, prompting a search for novel compounds with mechanisms of action that are different from the traditionally used drugs. The immunosuppressive drug FK506 shows an antimalarial activity. The mechanism of the drug action involves the molecular interaction with the parasite target proteins PfFKBP35 and PvFKBP35, which are novel...
Probing local structural fluctuations in myoglobin by size‐dependent thiol‐disulfide exchange
- Margaret M. Stratton, Thomas A. Cutler, Jeung‐Hoi Ha, Stewart N. Loh
- Published in Wiley Interscience on Jun 02, 2010
- DOI: 10.1002/pro.440 (p 1587-1594)
Abstract All proteins undergo local structural fluctuations (LSFs) or breathing motions. These motions are likely to be important for function but are poorly understood. LSFs were initially defined by amide hydrogen exchange (HX) experiments as opening events, which expose a small number of backbone amides to 1H/2H exchange, but whose exchange rates are independent of denaturant concentration. Here, we use size‐dependent thiol‐disulfide ...




