Electrostatic steering and ionic tethering in enzyme–ligand binding: Insights from simulations

To bind at an enzyme’s active site, a ligand must diffuse or be transported to the enzyme’s surface, and, if the binding site is buried, the ligand must diffuse through the protein to reach it. Although the driving force for ligand binding is often ascribed to the hydrophobic effect, electrostatic interactions also influence the binding process of both charged and nonpolar ligands. First, electrostatic steering of charged substrates into enzyme active sites is discussed. This is of particular relevance for diffusion-influenced enzymes. By comparing the results of Brownian dynamics simulations and electrostatic potential similarity analysis for triose-phosphate isomerases, superoxide dismutases, and β-lactamases from different species, we identify the conserved features responsible for the electrostatic substrate-steering fields. The conserved potentials are localized at the active sites and are the primary determinants of the bimolecular association rates. Then we focus on a more subtle effect, which we will refer to as “ionic tethering.” We explore, by means of molecular and Brownian dynamics simulations and electrostatic continuum calculations, how salt links can act as tethers between structural elements of an enzyme that undergo conformational change upon substrate binding, and thereby regulate or modulate substrate binding. This is illustrated for the lipase and cytochrome P450 enzymes. Ionic tethering can provide a control mechanism for substrate binding that is sensitive to the electrostatic properties of the enzyme’s surroundings even when the substrate is nonpolar.

Functional testicular tissue does not masculinize development of the zebra finch song system.

Current theories of sexual differentiation maintain that ovarian estrogen prevents masculine development of the copulatory system in birds, whereas estrogen derived from testicular androgens promotes masculine sexual differentiation of neuroanatomy and sexual behavior in mammals. Paradoxically, some data suggest that the neural song system in zebra finches follows the mammalian pattern with estrogenic metabolites of testicular secretions causing masculine development. To test whether the removal of estrogen from males during early development would prevent the development of masculine song systems, zebra finches were treated embryonically with an inhibitor of estrogen synthesis. In addition, this treatment in genetic female zebra finches induced both functional ovarian and testicular tissue to develop, thus allowing the assessment of the direct effects of testicular secretions on song system development. In males, the inhibition of estrogen synthesis before hatching had a small but significant effect in demasculinizing one aspect of the neural song system. In treated females, the song systems remained morphologically feminine. These results suggest that masculinization of the song system is not determined solely by testicular androgens or their estrogenic metabolites.

Solution structure of the Shc SH2 domain complexed with a tyrosine-phosphorylated peptide from the T-cell receptor.

She is a widely expressed adapter protein that plays an important role in signaling via a variety of cell surface receptors and has been implicated in coupling the stimulation of growth factor, cytokine, and antigen receptors to the Ras signaling pathway. She interacts with several tyrosine-phosphorylated receptors through its C-terminal SH2 domain, and one of the mechanisms of T-cell receptor-mediated Ras activation involves the interaction of the Shc SH2 domain with the tyrosine-phosphorylated zeta chain of the T-cell receptor. Here we describe a high-resolution NMR structure of the Shc SH2 domain complexed to a phosphopeptide (GHDGLpYQGLSTATK) corresponding to a portion of the zeta chain of the T-cell receptor. Although the overall architecture of the protein is similar to other SH2 domains, distinct structural differences were observed in the smaller beta-sheet, BG loop, (pY + 3) phosphopeptide-binding site, and relative position of the bound phosphopeptide.

Delineation of a slow-twitch-myofiber-specific transcriptional element by using in vivo somatic gene transfer.

Contractile proteins are encoded by multigene families, most of whose members are differentially expressed in fast- versus slow-twitch myofibers. This fiber-type-specific gene regulation occurs by unknown mechanisms and does not occur within cultured myocytes. We have developed a transient, whole-animal assay using somatic gene transfer to study this phenomenon and have identified a fiber-type-specific regulatory element within the promoter region of a slow myofiber-specific gene. A plasmid-borne luciferase reporter gene fused to various muscle-specific contractile gene promoters was differentially expressed when injected into slow- versus fast-twitch rat muscle: the luciferase gene was preferentially expressed in slow muscle when fused to a slow troponin I promoter, and conversely, was preferentially expressed in fast muscle when fused to a fast troponin C promoter. In contrast, the luciferase gene was equally well expressed by both muscle types when fused to a nonfiber-type-specific skeletal actin promoter. Deletion analysis of the troponin I promoter region revealed that a 157-bp enhancer conferred slow-muscle-preferential activity upon a minimal thymidine kinase promoter. Transgenic analysis confirmed the role of this enhancer in restricting gene expression to slow-twitch myofibers. Hence, somatic gene transfer may be used to rapidly define elements that direct myofiber-type-specific gene expression prior to the generation of transgenic mice.

Library of libraries: approach to synthetic combinatorial library design and screening of "pharmacophore" motifs.

Construction of synthetic combinatorial libraries is described that allows for the generation of a library of motifs rather than a library of compounds. Peptide libraries based on this strategy were synthesized and screened with model targets streptavidin and anti-beta-endorphin antibody. The screens resulted in observation of expected motifs providing evidence of the effectiveness of the suggested approach.

Asking For Help: Vulnerability within Consultative Assessment A Couples Therapy Example.

In many different ways psychologists are challenged to evaluate their work and be accountable when therapy is not progressing. One time honored approach to addressing the lack of progress is to seek formal psychological assessment to gain a better diagnostic understanding as well as treatment recommendations. The perspective on the purpose of the assessment seems to have changed however. Historically, the presumption was that there was some kind of pathology that the clinician was not seeing, where currently the perspective seems to be shifting more towards a focus on what might not be working in the therapeutic process. This shift in focus has led to new styles of collaborative and consultative assessment such as Therapeutic Assessment. Therapeutic Assessment was developed by Stephen Finn and has shown a great deal of promise helping clients and therapists gain valuable insight and move toward meaningful life changes (1998). Amidst the promise of consultative assessment also lie potential challenges for the referring clinician. This paper will explore the nature of consultative relationships in the field of mental health and the current thoughts about the advantages and challenges of these relationships. Using a case study format, it will discuss and illustrate issues of professional vulnerability that can arise during these relationships and postulate that a similar vulnerability could be present during a consultative assessment. Recommendations are made for maximizing the success of this type of assessment, and ideas for future research are reviewed. Through the lens of a couples Therapeutic Assessment case, this paper will examine what it is about this unique kind of professional relationship that garners such powerful potential and pitfalls.

Slaying the Hydra: Al Qaeda's Evolution and America's Plan of Attack

In response to 9/11, the U.S. launched airstrikes in Afghanistan against al Qaeda to diminish its ability to conduct future attacks. Next, the U.S. turned its attention to Iraq. A series of problems and missteps there subsequently created an opportunity for al Qaeda to evolve, aligning itself with active insurgencies and other terrorist groups around the world. This paper identifies the largely preemptive strategies employed by the U.S. and analyzes how those efforts positively and negatively impacted the War on Terror. While some significant gains were made, the results show that al Qaeda remains a threat to the international community and demands more effective U.S. and international counter-radicalization strategies, including diplomacy and aid, to curb its global appeal.

Rock map of the Yellow River (Huang He) expedition, China, 1925-1927 : Sheet 2

This layer is part of a set of georeferenced, raster images of the manuscript, paper map set entitled: Ch'ing-Hai upper Yellow River expedition : Rock and Simpson, 1925-27, [cartography by J.F. Rock]. Scale 1:250,000. This layer image is of Sheet II [of 10] covering a portion of the Yellow River (Huang He) region in eastern Qinghai Sheng and southern Gansu Sheng, China. The map set details the route and surrounding environs of the Arnold Arboretum's "Western China" expedition led by Joseph Rock, 1924-1927. The set covers a portion of the Yellow River (Huang He) region in south central China (Qinghai, Gansu, and Sichuan shengs (a portion of historic Tibet)). It shows features, labeled variously in English, Chinese, Wade-Giles transliteration, and Tibetan, including: rivers, streams, lakes, mountains, gorges, valleys, plateaus, plains, cities, towns, villages, provincial capitals, county seats, passes, monasteries, ruin sites, native tribe locations, and more. Relief is shown by hachures, spot heights, and landform drawings. The original manuscript map set is part of the Harvard College Library, Harvard Map Collection. "Joseph Rock traced his travels for the [Arnold] Arboretum's [Western China] 1924-1927 expedition in a colorful, hand-drawn map entitled 'Ch'ing-Hai upper Yellow River expedition.' The pen-and-ink drawing was made on ten sheets that when joined form a single, irregularly-shaped map, approximately six by eight feet in size. The individual sheets are numbered, using roman numerals; on sheet VII is a second title, 'Choni Territory, Upper and Lower T'ieh-Pu country and route to Sung-Pan, J. F. Rock, 1925-1927.' Topographical and other features are identified using a combination of English, Chinese characters, Wade-Giles transliterations and Tibetan script. Rock's attractive cursive style and use of hachures, spot heights, and landform drawings to depict relief add character to the map." -- Text from the Arnold Arboretum Web site.

Rock map of the Yellow River (Huang He) expedition, China, 1925-1927 : Sheet 3

This layer is part of a set of georeferenced, raster images of the manuscript, paper map set entitled: Ch'ing-Hai upper Yellow River expedition : Rock and Simpson, 1925-27, [cartography by J.F. Rock]. Scale 1:250,000. This layer image is of Sheet III [of 10] covering a portion of the Yellow River (Huang He) region in eastern Qinghai Sheng and southern Gansu Sheng, China. The map set details the route and surrounding environs of the Arnold Arboretum's "Western China" expedition led by Joseph Rock, 1924-1927. The set covers a portion of the Yellow River (Huang He) region in south central China (Qinghai, Gansu, and Sichuan shengs (a portion of historic Tibet)). It shows features, labeled variously in English, Chinese, Wade-Giles transliteration, and Tibetan, including: rivers, streams, lakes, mountains, gorges, valleys, plateaus, plains, cities, towns, villages, provincial capitals, county seats, passes, monasteries, ruin sites, native tribe locations, and more. Relief is shown by hachures, spot heights, and landform drawings. The original manuscript map set is part of the Harvard College Library, Harvard Map Collection. "Joseph Rock traced his travels for the [Arnold] Arboretum's [Western China] 1924-1927 expedition in a colorful, hand-drawn map entitled 'Ch'ing-Hai upper Yellow River expedition.' The pen-and-ink drawing was made on ten sheets that when joined form a single, irregularly-shaped map, approximately six by eight feet in size. The individual sheets are numbered, using roman numerals; on sheet VII is a second title, 'Choni Territory, Upper and Lower T'ieh-Pu country and route to Sung-Pan, J. F. Rock, 1925-1927.' Topographical and other features are identified using a combination of English, Chinese characters, Wade-Giles transliterations and Tibetan script. Rock's attractive cursive style and use of hachures, spot heights, and landform drawings to depict relief add character to the map." -- Text from the Arnold Arboretum Web site.

Rock map of the Yellow River (Huang He) expedition, China, 1925-1927 : Sheet 4

This layer is part of a set of georeferenced, raster images of the manuscript, paper map set entitled: Ch'ing-Hai upper Yellow River expedition : Rock and Simpson, 1925-27, [cartography by J.F. Rock]. Scale 1:250,000. This layer image is of Sheet IV [of 10] covering a portion of the Yellow River (Huang He) region in eastern Qinghai Sheng, China. The map set details the route and surrounding environs of the Arnold Arboretum's "Western China" expedition led by Joseph Rock, 1924-1927. The set covers a portion of the Yellow River (Huang He) region in south central China (Qinghai, Gansu, and Sichuan shengs (a portion of historic Tibet)). It shows features, labeled variously in English, Chinese, Wade-Giles transliteration, and Tibetan, including: rivers, streams, lakes, mountains, gorges, valleys, plateaus, plains, cities, towns, villages, provincial capitals, county seats, passes, monasteries, ruin sites, native tribe locations, and more. Relief is shown by hachures, spot heights, and landform drawings. The original manuscript map set is part of the Harvard College Library, Harvard Map Collection. "Joseph Rock traced his travels for the [Arnold] Arboretum's [Western China] 1924-1927 expedition in a colorful, hand-drawn map entitled 'Ch'ing-Hai upper Yellow River expedition.' The pen-and-ink drawing was made on ten sheets that when joined form a single, irregularly-shaped map, approximately six by eight feet in size. The individual sheets are numbered, using roman numerals; on sheet VII is a second title, 'Choni Territory, Upper and Lower T'ieh-Pu country and route to Sung-Pan, J. F. Rock, 1925-1927.' Topographical and other features are identified using a combination of English, Chinese characters, Wade-Giles transliterations and Tibetan script. Rock's attractive cursive style and use of hachures, spot heights, and landform drawings to depict relief add character to the map." -- Text from the Arnold Arboretum Web site.

Rock map of the Yellow River (Huang He) expedition, China, 1925-1927 : Sheet 9

This layer is part of a set of georeferenced, raster images of the manuscript, paper map set entitled: Ch'ing-Hai upper Yellow River expedition : Rock and Simpson, 1925-27, [cartography by J.F. Rock]. Scale 1:250,000. This layer image is of Sheet IX [of 10] covering a portion of the Yellow River (Huang He) region in southern Gansu Sheng and northwestern Sichuan Sheng, China, including parts of Baishui Jiang and Pai Ho (Gar He). The map set details the route and surrounding environs of the Arnold Arboretum's "Western China" expedition led by Joseph Rock, 1924-1927. The set covers a portion of the Yellow River (Huang He) region in south central China (Qinghai, Gansu, and Sichuan shengs (a portion of historic Tibet)). It shows features, labeled variously in English, Chinese, Wade-Giles transliteration, and Tibetan, including: rivers, streams, lakes, mountains, gorges, valleys, plateaus, plains, cities, towns, villages, provincial capitals, county seats, passes, monasteries, ruin sites, native tribe locations, and more. Relief is shown by hachures, spot heights, and landform drawings. The original manuscript map set is part of the Harvard College Library, Harvard Map Collection. "Joseph Rock traced his travels for the [Arnold] Arboretum's [Western China] 1924-1927 expedition in a colorful, hand-drawn map entitled 'Ch'ing-Hai upper Yellow River expedition.' The pen-and-ink drawing was made on ten sheets that when joined form a single, irregularly-shaped map, approximately six by eight feet in size. The individual sheets are numbered, using roman numerals; on sheet VII is a second title, 'Choni Territory, Upper and Lower T'ieh-Pu country and route to Sung-Pan, J. F. Rock, 1925-1927.' Topographical and other features are identified using a combination of English, Chinese characters, Wade-Giles transliterations and Tibetan script. Rock's attractive cursive style and use of hachures, spot heights, and landform drawings to depict relief add character to the map." -- Text from the Arnold Arboretum Web site.

Rock map of the Yellow River (Huang He) expedition, China, 1925-1927 : Sheet 5

This layer is part of a set of georeferenced, raster images of the manuscript, paper map set entitled: Ch'ing-Hai upper Yellow River expedition : Rock and Simpson, 1925-27, [cartography by J.F. Rock]. Scale 1:250,000. This layer image is of Sheet V [of 10] covering a portion of the Yellow River (Huang He) region in eastern Qinghai Sheng and southern Gansu Sheng, China. The map set details the route and surrounding environs of the Arnold Arboretum's "Western China" expedition led by Joseph Rock, 1924-1927. The set covers a portion of the Yellow River (Huang He) region in south central China (Qinghai, Gansu, and Sichuan shengs (a portion of historic Tibet)). It shows features, labeled variously in English, Chinese, Wade-Giles transliteration, and Tibetan, including: rivers, streams, lakes, mountains, gorges, valleys, plateaus, plains, cities, towns, villages, provincial capitals, county seats, passes, monasteries, ruin sites, native tribe locations, and more. Relief is shown by hachures, spot heights, and landform drawings. The original manuscript map set is part of the Harvard College Library, Harvard Map Collection. "Joseph Rock traced his travels for the [Arnold] Arboretum's [Western China] 1924-1927 expedition in a colorful, hand-drawn map entitled 'Ch'ing-Hai upper Yellow River expedition.' The pen-and-ink drawing was made on ten sheets that when joined form a single, irregularly-shaped map, approximately six by eight feet in size. The individual sheets are numbered, using roman numerals; on sheet VII is a second title, 'Choni Territory, Upper and Lower T'ieh-Pu country and route to Sung-Pan, J. F. Rock, 1925-1927.' Topographical and other features are identified using a combination of English, Chinese characters, Wade-Giles transliterations and Tibetan script. Rock's attractive cursive style and use of hachures, spot heights, and landform drawings to depict relief add character to the map." -- Text from the Arnold Arboretum Web site.

Rock map of the Yellow River (Huang He) expedition, China, 1925-1927 : Sheet 6

This layer is part of a set of georeferenced, raster images of the manuscript, paper map set entitled: Ch'ing-Hai upper Yellow River expedition : Rock and Simpson, 1925-27, [cartography by J.F. Rock]. Scale 1:250,000. This layer image is of Sheet VI [of 10] covering a portion of the Yellow River (Huang He) region in eastern Qinghai Sheng, southern Gansu Sheng, and northwestern Sichuan Sheng, China. The map set details the route and surrounding environs of the Arnold Arboretum's "Western China" expedition led by Joseph Rock, 1924-1927. The set covers a portion of the Yellow River (Huang He) region in south central China (Qinghai, Gansu, and Sichuan shengs (a portion of historic Tibet)). It shows features, labeled variously in English, Chinese, Wade-Giles transliteration, and Tibetan, including: rivers, streams, lakes, mountains, gorges, valleys, plateaus, plains, cities, towns, villages, provincial capitals, county seats, passes, monasteries, ruin sites, native tribe locations, and more. Relief is shown by hachures, spot heights, and landform drawings. The original manuscript map set is part of the Harvard College Library, Harvard Map Collection. "Joseph Rock traced his travels for the [Arnold] Arboretum's [Western China] 1924-1927 expedition in a colorful, hand-drawn map entitled 'Ch'ing-Hai upper Yellow River expedition.' The pen-and-ink drawing was made on ten sheets that when joined form a single, irregularly-shaped map, approximately six by eight feet in size. The individual sheets are numbered, using roman numerals; on sheet VII is a second title, 'Choni Territory, Upper and Lower T'ieh-Pu country and route to Sung-Pan, J. F. Rock, 1925-1927.' Topographical and other features are identified using a combination of English, Chinese characters, Wade-Giles transliterations and Tibetan script. Rock's attractive cursive style and use of hachures, spot heights, and landform drawings to depict relief add character to the map." -- Text from the Arnold Arboretum Web site.