Faculty and Staff

A structurally and compositionally well-defined and spectrally tunable artificial light-harvesting system has been constructed in which multiple organic dyes attached to a three-arm-DNA nanostructure serve as an antenna conjugated to a photosynthetic reaction center isolated from Rhodobacter sphaeroides 2.4.1. The light energy absorbed by the dye molecules is transferred to the reaction center, where charge separation takes place. The average number of DNA three-arm junctions per reaction center was tuned from 0.75 to 2.35. This DNA-templated multichromophore system serves as a modular light-harvesting antenna that is capable of being optimized for its spectral properties, energy transfer efficiency, and photostability, allowing one to adjust both the size and spectrum of the resulting structures. This may serve as a useful test bed for developing nanostructured photonic systems.

Time-resolved fluorescence spectroscopy was used to explore the pathway and kinetics of energy transfer in photosynthetic membrane vesicles (chromatophores) isolated from Rhodobacter (Rba.) sphaeroides cells harvested 2, 4, 6 or 24 hours after a transition from growth in high to low level illumination. As previously observed, this light intensity transition initiates the remodeling of the photosynthetic apparatus and an increase in the number of light harvesting 2 (LH2) complexes relative to light harvesting 1 (LH1) and reaction center (RC) complexes. It has generally been thought that the increase in LH2 complexes served the purpose of increasing the overall energy transmission to the RC. However, fluorescence lifetime measurements and analysis in terms of energy transfer within LH2 and between LH2 and LH1 indicate that, during the remodeling time period measured, only a portion of the additional LH2 generated are well connected to LH1 and the reaction center. The majority of the additional LH2 fluorescence decays with a lifetime comparable to that of free, unconnected LH2 complexes. The presence of large LH2-only domains has been observed by atomic force microscopy in Rba. sphaeroides chromatophores (Bahatyrova et al., Nature, 2004, 430, 1058), providing structural support for the existence of pools of partially connected LH2 complexes. These LH2-only domains represent the light-responsive antenna complement formed after a switch in growth conditions from high to low illumination, while the remaining LH2 complexes occupy membrane regions containing mixtures of LH2 and LH1–RC core complexes. The current study utilized a multi-parameter approach to explore the fluorescence spectroscopic properties related to the remodeling process, shedding light on the structure-function relationship of the photosynthetic assembles. Possible reasons for the accumulation of these largely disconnected LH2-only pools are discussed.

In 2014/2015, Arizona State University (ASU) Libraries, the Labriola National American Indian Data Center, and the ASU American Indian Studies Department completed an ASU Institute for Humanities Research (IHR) seed grant entitled “Carlos Montezuma’s Wassaja Newsletter: Digitization, Access and Context” to digitize all ASU held issues of the newsletter Wassaja Freedom’s Signal for the Indian, which Yavapai activist-intellectual Carlos Montezuma, MD (1866-1923) self-published during 1916-1922. The grant team additionally selected a portion of the ASU Libraries Carlos Montezuma archival collection for digitization to provide a more complete picture of Dr. Carlos Montezuma’s life and work.

The ASU grant team produced a searchable online collection on the ASU Digital Repository and created an online exhibition in conjunction with the IHR Nexus Lab’s Developing Wassaja Project. The Nexus Lab’s role at ASU is to grow the digital humanities through interdisciplinary collaborations bringing together humanities, science, and technology. The Nexus Lab partnered with the grant team to create the Developing Wassaja Project which provided an opportunity for faculty, staff, and students at ASU to engage in electronic publication through web application development.

The resulting web platform, Wassaja: A Carlos Montezuma Project, provides context for this digitized collection and facilitates community interaction, including a partnership with Dr. Montezuma’s home community the Fort McDowell Yavapai Nation. In this webcast, Digital Projects Librarian Matthew Harp, Developing Wassaja Project team member Joe Buenker (subject librarian), and grant team member Joyce Martin (librarian and curator of the Labriola National American Indian Data Center) will discuss and demonstrate the resources created and the resulting partnership with the Fort McDowell Yavapai Nation. The webcast will focus on identifying collaborators and needed skills to engage in Digital Humanities research and on identifying the stages of a collaborative project.

Participants will gain insight on working directly with diverse communities; overcoming technical limitations of traditional institutional repositories; collaborative strategies with faculty, research centers, and cultural heritage societies; solutions for moving hidden collections into an engaging digital exhibition; integrating digital humanities research and instruction with library curation; and preparing for long term costs and management issues.

Anthropology librarian Juliann Couture and Joyce Martin, curator of the Labriola National American Indian Data Center, looking at the Center's display of unique Hopi Kachina dolls. Four of the kachinas (Navan Kachina; Talavi Kachina; Flute Kachina; and Ahöla Kachina) were created by artist, carver, and former ASU employee Tony Dukepoo as a gift to the libraries in 1979. The kachina dolls are on display in the Labriola Center located on the 2nd floor of the Hayden Library on ASU's Tempe campus.

The Simon Ortiz and Labriola Center Lecture on Indigenous Land, Culture, and Community addresses topics and issues across disciplines in the arts, humanities, sciences, and politics. Underscoring Indigenous American experiences and perspectives, this Series seeks to create and celebrate knowledge that evolves from an Indigenous worldview that is inclusive and that is applicable to all walks of life.” Professor Simon Ortiz discussed the overall nature of the Series, especially emphasizing the global nature of Indigenous concerns. Joyce Martin and Matthew Harp elaborated on the contributions of the Labriola National American Indian Data Center and ASU Libraries to the Series.

The Labriola Center hosts an informal event in Hayden Library which facilitates close interaction between the featured speaker and audience members. The ASU Libraries records the evening lectures which take place at the Heard Museum and presents both an audio podcast and streaming video of each lecture on the ASU Library Channel webpage. This lecture series provides not only a chance for community discussion at the events themselves, but through the innovative use of technology the ASU Libraries enables additional forums for discussion in blogs and web pages which choose to link to the streaming videos.

There are an increasing variety of applications in which peptides are both synthesized and used attached to solid surfaces. This has created a need for high throughput sequence analysis directly on surfaces. However, common sequencing approaches that can be adapted to surface bound peptides lack the throughput often needed in library-based applications. Here we describe a simple approach for sequence analysis directly on solid surfaces that is both high speed and high throughput, utilizing equipment available in most protein analysis facilities. In this approach, surface bound peptides, selectively labeled at their N-termini with a positive charge-bearing group, are subjected to controlled degradation in ammonia gas, resulting in a set of fragments differing by a single amino acid that remain spatially confined on the surface they were bound to. These fragments can then be analyzed by MALDI mass spectrometry, and the peptide sequences read directly from the resulting spectra.

Background: Chemistry and particularly enzymology at surfaces is a topic of rapidly growing interest, both in terms of its role in biological systems and its application in biocatalysis. Existing protein immobilization approaches, including noncovalent or covalent attachments to solid supports, have difficulties in controlling protein orientation, reducing nonspecific absorption and preventing protein denaturation. New strategies for enzyme immobilization are needed that allow the precise control over orientation and position and thereby provide optimized activity.

Methodology/Principal Findings: A method is presented for utilizing peptide ligands to immobilize enzymes on surfaces with improved enzyme activity and stability. The appropriate peptide ligands have been rapidly selected from high-density arrays and when desirable, the peptide sequences were further optimized by single-point variant screening to enhance both the affinity and activity of the bound enzyme. For proof of concept, the peptides that bound to β-galactosidase and optimized its activity were covalently attached to surfaces for the purpose of capturing target enzymes. Compared to conventional methods, enzymes immobilized on peptide-modified surfaces exhibited higher specific activity and stability, as well as controlled protein orientation.

Conclusions/Significance: A simple method for immobilizing enzymes through specific interactions with peptides anchored on surfaces has been developed. This approach will be applicable to the immobilization of a wide variety of enzymes on surfaces with optimized orientation, location and performance, and provides a potential mechanism for the patterned self-assembly of multiple enzymes on surfaces.

Cells routinely compartmentalize enzymes for enhanced efficiency of their metabolic pathways. Here we report a general approach to construct DNA nanocaged enzymes for enhancing catalytic activity and stability. Nanocaged enzymes are realized by self-assembly into DNA nanocages with well-controlled stoichiometry and architecture that enabled a systematic study of the impact of both encapsulation and proximal polyanionic surfaces on a set of common metabolic enzymes. Activity assays at both bulk and single-molecule levels demonstrate increased substrate turnover numbers for DNA nanocage-encapsulated enzymes. Unexpectedly, we observe a significant inverse correlation between the size of a protein and its activity enhancement. This effect is consistent with a model wherein distal polyanionic surfaces of the nanocage enhance the stability of active enzyme conformations through the action of a strongly bound hydration layer. We further show that DNA nanocages protect encapsulated enzymes against proteases, demonstrating their practical utility in functional biomaterials and biotechnology.