Journal Issue - Volume 18 Issue 11 (November 2009)
In this issue
- Published in Wiley Interscience on Oct 23, 2009
- DOI: 10.1002/pro.271 (p n/a-n/a)
The Drosophila G protein‐coupled receptor, Methuselah, exhibits a promiscuous response to peptides
- William W. Ja, Gil B. Carvalho, Marisol Madrigal, Richard W. Roberts, Seymour Benzer
- Published in Wiley Interscience on Aug 11, 2009
- DOI: 10.1002/pro.221 (p 2203-2208)
Abstract Methuselah (Mth) is a G protein‐coupled receptor (GPCR) associated with longevity in Drosophila melanogaster. Previously, Stunted (Sun) was identified as a peptide agonist of Mth. Here, we identify two additional activators of Mth signaling: Drosophila Sex Peptide (SP) and a novel peptide (Serendipitous Peptide Activator of Mth, SPAM). Minimal functional sequences and key residues were identified from Sun and SPAM by studying...
Elucidation of the stability and functional regions of the human coronavirus OC43 nucleocapsid protein
- Chun‐Yu Huang, Yen‐Lan Hsu, Wan‐Ling Chiang, Ming‐Hon Hou
- Published in Wiley Interscience on Aug 18, 2009
- DOI: 10.1002/pro.225 (p 2209-2218)
Abstract Human coronavirus OC43 (HCoV‐OC43) is one of the causes of the “common cold” in human during seasons of cold weather. The primary function of the HCoV‐OC43 nucleocapsid protein (N protein) is to recognize viral genomic RNA, which leads to ribonucleocapsid formation. Here, we characterized the stability and identified the functional regions of the recombinant HCoV‐OC43 N protein. Circular dichroism and fluorescence...
Designed zinc finger protein interacting with the HIV‐1 integrase recognition sequence at 2‐LTR‐circle junctions
- Supachai Sakkhachornphop, Supat Jiranusornkul, Kanchanok Kodchakorn, Sawitree Nangola, Thira Sirisanthana, Chatchai Tayapiwatana
- Published in Wiley Interscience on Aug 21, 2009
- DOI: 10.1002/pro.233 (p 2219-2230)
Abstract Integration of HIV‐1 cDNA into the host genome is a crucial step for viral propagation. Two nucleotides, cytosine and adenine (CA), conserved at the 3′ end of the viral cDNA genome, are cleaved by the viral integrase (IN) enzyme. As IN plays a crucial role in the early stages of the HIV‐1 life cycle, substrate blockage of IN is an attractive strategy for therapeutic interference. In this study, we used the 2‐LTR‐circle...
A kinetic assessment of the C. elegans amyloid disaggregation activity enables uncoupling of disassembly and proteolysis
- Jan Bieschke, Ehud Cohen, Amber Murray, Andrew Dillin, Jeffery W. Kelly
- Published in Wiley Interscience on Aug 21, 2009
- DOI: 10.1002/pro.234 (p 2231-2241)
Abstract Protein aggregation is a common feature of late onset neurodegenerative disorders, including Alzheimer's disease. In Alzheimer's disease, misassembly of the Aβ peptide is genetically linked to proteotoxicity associated with disease etiology. A reduction in Aβ proteotoxicity is accomplished, in part, by the previously reported Aβ disaggregation and proteolysis activities–under partial control of heat shock factor 1, a...
Human lysosomal α‐mannosidases exhibit different inhibition and metal binding properties
- Meenakshi Venkatesan, Douglas A. Kuntz, David R. Rose
- Published in Wiley Interscience on Aug 31, 2009
- DOI: 10.1002/pro.235 (p 2242-2251)
Abstract Two structurally‐related members of the lysosomal mannosidase family, the broad substrate specificity enzyme human lysosomal α‐mannosidase (hLM, MAN2B1) and the human core α‐1, 6‐specific mannosidase (hEpman, MAN2B2) act in a complementary fashion on different glycosidic linkages, to effect glycan degradation in the lysosome. We have successfully expressed these enzymes in Drosophila S2 cells and functionally characterized...
Structure of the Cdt1 C‐terminal domain: Conservation of the winged helix fold in replication licensing factors
- Bulat I. Khayrutdinov, Won Jin Bae, Young Mi Yun, Jie Hye Lee, Takashi Tsuyama, Jung Joo Kim, Eunha Hwang, Kyoung‐Seok Ryu, Hae‐Kap Cheong, Chaejoon Cheong, Jung‐Soon Ko, Takemi Enomoto, P. Andrew Karplus, Peter Güntert, Shusuke Tada, Young Ho Jeon, Yunje Cho
- Published in Wiley Interscience on Aug 31, 2009
- DOI: 10.1002/pro.236 (p 2252-2264)
Abstract In eukaryotic replication licensing, Cdt1 plays a key role by recruiting the MCM2‐7 complex onto the origin of chromosome. The C‐terminal domain of mouse Cdt1 (mCdt1C), the most conserved region in Cdt1, is essential for licensing and directly interacts with the MCM2‐7 complex. We have determined the structures of mCdt1CS (mCdt1C_small; residues 452 to 557) and mCdt1CL (mCdt1C_large; residues 420 to 557) using X‐ray...
Structure and interaction of ubiquitin‐associated domain of human Fas‐associated factor 1
- JinSue Song, Joon Kyu Park, Jae‐Jin Lee, Yun‐Seok Choi, Kyoung‐Seok Ryu, Jae‐Hong Kim, Eunhee Kim, Kong‐Joo Lee, Young‐Ho Jeon, Eunice EunKyeong Kim
- Published in Wiley Interscience on Aug 31, 2009
- DOI: 10.1002/pro.237 (p 2265-2276)
Abstract Fas‐associated factor (FAF)‐1 is a multidomain protein that was first identified as a member of the Fas death‐inducing signaling complex, but later found to be involved in various biological processes. Although the exact mechanisms are not clear, FAF1 seems to play an important role in cancer, asbestos‐induced mesotheliomas, and Parkinson's disease. It interacts with polyubiquitinated proteins, Hsp70, and p97/VCP...
Domain 4 of the anthrax protective antigen maintains structure and binding to the host receptor CMG2 at low pH
- Alexander S. Williams, Scott Lovell, Asokan Anbanandam, Rahif El‐Chami, James G. Bann
- Published in Wiley Interscience on Aug 31, 2009
- DOI: 10.1002/pro.238 (p 2277-2286)
Abstract Domain 4 of the anthrax protective antigen (PA) plays a key role in cellular receptor recognition as well as in pH‐dependent pore formation. We present here the 1.95 Å crystal structure of domain 4, which adopts a fold that is identical to that observed in the full‐length protein. We have also investigated the structural properties of the isolated domain 4 as a function of pH, as well as the pH‐dependence on binding to the...
The interaction of Bacillus subtilis σ A with RNA polymerase
- Elecia B. Johnston, Peter J. Lewis, Renate Griffith
- Published in Wiley Interscience on Sep 04, 2009
- DOI: 10.1002/pro.239 (p 2287-2297)
Abstract RNA polymerase (RNAP) is an essential and highly conserved enzyme in all organisms. The process of transcription initiation is fundamentally different between prokaryotes and eukaryotes. In prokaryotes, initiation is regulated by σ factors, making the essential interaction between σ factors and RNAP an attractive target for antimicrobial agents. Our objective was to achieve the first step in the process of developing novel...
Role of transmembrane domain 10 for the function of organic anion transporting polypeptide 1B1
- Chunshan Gui, Bruno Hagenbuch
- Published in Wiley Interscience on Sep 16, 2009
- DOI: 10.1002/pro.240 (p 2298-2306)
Abstract The liver‐specific organic anion transporting polypeptides OATP1B1 and OATP1B3 are highly homologous and share numerous substrates. However, at low concentrations OATP1B1 shows substrate selectivity for estrone‐3‐sulfate. In this study, we investigated the molecular mechanism for this substrate selectivity of OATP1B1 by constructing OATP1B1/1B3 chimeric transporters and by site‐directed mutagenesis. Functional studies of...
The oligomerization of CynR in Escherichia coli
- Gwendowlyn S. Knapp, James C. Hu
- Published in Wiley Interscience on Sep 16, 2009
- DOI: 10.1002/pro.241 (p 2307-2315)
Abstract Deletion analysis and alanine‐scanning based on a homology‐based interaction model were used to identify determinants of oligomerization in the transcriptional regulator CynR, a member of the LysR‐type transcriptional regulator (LTTR) family. Deletion analysis confirmed that the putative regulatory domain of CynR was essential for driving the oligomerization of λ repressor‐CynR fusion proteins. The interaction surface of a...
One‐plasmid tunable coexpression for mycobacterial protein–protein interaction studies
- Yong Chang, David Mead, Vinay Dhodda, Phil Brumm, Brian G. Fox
- Published in Wiley Interscience on Sep 16, 2009
- DOI: 10.1002/pro.242 (p 2316-2325)
Abstract A single plasmid that allows controlled coexpression has been developed for use in mycobacteria. The tetracycline inducible promoter, PtetO, was used to provide tetracycline‐dependent induction of one gene, while the Psmyc, Pimyc, or Phsp promoters were used to provide three different levels of constitutive expression of a second gene. The functions of these four individual promoters were established using green fluorescent...
Trp fluorescence reveals an activation‐dependent cation‐π interaction in the Switch II region of Gα i proteins
- Heidi E. Hamm, Scott M. Meier, Guihua Liao, Anita M. Preininger
- Published in Wiley Interscience on Sep 16, 2009
- DOI: 10.1002/pro.243 (p 2326-2335)
Abstract Crystal structures of Gαi (and closely related family member Gαt) reveal much of what we currently know about G protein structure, including changes which occur in Switch regions. Gαt exhibits a low rate of basal (uncatalyzed) nucleotide exchange and an ordered Switch II region in the GDP‐bound state, unlike Gαi, which exhibits higher basal exchange and a disordered Switch II region in GαiGDP structures. Using purified Gαi and Gαt, we examined the...
Crystal structures of a therapeutic single chain antibody in complex with two drugs of abuse—Methamphetamine and 3,4‐methylenedioxymethamphetamine
- Reha Celikel, Eric C. Peterson, S. Michael Owens, Kottayil I. Varughese
- Published in Wiley Interscience on Sep 16, 2009
- DOI: 10.1002/pro.244 (p 2336-2345)
Abstract Methamphetamine (METH) is a major drug threat in the United States and worldwide. Monoclonal antibody (mAb) therapy for treating METH abuse is showing exciting promise and the understanding of how mAb structure relates to function will be essential for future development of these important therapies. We have determined crystal structures of a high affinity anti‐(+)‐METH therapeutic single chain antibody fragment (scFv6H4,...




