ASU Scholarship Showcase

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.

Background: Little research has explored who responds better to an automated vs. human advisor for health behaviors in general, and for physical activity (PA) promotion in particular. The purpose of this study was to explore baseline factors (i.e., demographics, motivation, interpersonal style, and external resources) that moderate intervention efficacy delivered by either a human or automated advisor.

Methods: Data were from the CHAT Trial, a 12-month randomized controlled trial to increase PA among underactive older adults (full trial N = 218) via a human advisor or automated interactive voice response advisor. Trial results indicated significant increases in PA in both interventions by 12 months that were maintained at 18-months. Regression was used to explore moderation of the two interventions.

Results: Results indicated amotivation (i.e., lack of intent in PA) moderated 12-month PA (d = 0.55, p < 0.01) and private self-consciousness (i.e., tendency to attune to one’s own inner thoughts and emotions) moderated 18-month PA (d = 0.34, p < 0.05) but a variety of other factors (e.g., demographics) did not (p > 0.12).

Conclusions: Results provide preliminary evidence for generating hypotheses about pathways for supporting later clinical decision-making with regard to the use of either human- vs. computer-delivered interventions for PA promotion.

The relationship between sequence and binding properties of an aptamer for immunoglobulin E (IgE) was investigated using custom DNA microarrays. Single, double and some triple mutations of the aptamer sequence were created to evaluate the importance of specific base composition on aptamer binding. The majority of the positions in the aptamer sequence were found to be immutable, with changes at these positions resulting in more than a 100-fold decrease in binding affinity. Improvements in binding were observed by altering the stem region of the aptamer, suggesting that it plays a significant role in binding. Results obtained for the various mutations were used to estimate the information content and the probability of finding a functional aptamer sequence by selection from a random library. For the IgE-binding aptamer, this probability is on the order of 10^-10 to 10^-9. Results obtained for the double and triple mutations also show that there are no compensatory mutations within the space defined by those mutations. Apparently, at least for this particular aptamer, the functional sequence space can be represented as a rugged landscape with sharp peaks defined by highly constrained base compositions. This makes the rational optimization of aptamer sequences using step-wise mutagenesis approaches very challenging.

Background: To identify social ecological correlates of objectively measured workplace sedentary behavior.

Methods: Participants from 24 worksites - across academic, industrial, and government sectors - wore an activPAL-micro accelerometer for 7-days (Jan-Nov 2016). Work time was segmented using daily logs. Sedentary behavior outcomes included time spent sitting, standing, in light intensity physical activity (LPA, stepping cadence <100 steps/min), and in prolonged sitting bouts (>30 min). Outcomes were standardized to an 8 h work day. Two electronic surveys were completed to derive individual (job type and work engagement), cultural (lunch away from the desk, walking at lunch and face-to-face interaction), physical (personal printer and office type) and organizational (sector) factors. Mixed-model analyses with worksite-level clustering were performed to examine multi-level associations. Secondary analyses examined job type and sector as moderators of these associations. All models were adjusted for age, race/ethnicity and gender.

Results: Participants (N = 478; 72% female; age: 45.0 ± 11.3 years; 77.8% non-Hispanic white) wore the activPAL-micro for 90.2 ± 15.5% of the reported workday. Walking at lunch was positively associated with LPA (5.0 ± 0.5 min/8 h, P < 0.001). Regular face-to-face interaction was negatively associated with prolonged sitting (−11.3 ± 4.8 min/8 h, P < 0.05). Individuals in private offices sat more (20.1 ± 9.1 min/8 h, P < 0.05), stood less (−21.5 ± 8.8 min/8 h, P < 0.05), and engaged in more prolonged sitting (40.9 ± 11.2 min/8 h, P < 0.001) than those in public office space. These associations were further modified by job type and sector.

Conclusions: Work-specific individual, cultural, physical and organizational factors are associated with workplace sedentary behavior. Associations vary by job type and sector and should be considered in the design of workplace interventions to reduce sedentary behavior.

Background: Although current technological advancements have allowed for objective measurements of sedentary behavior via accelerometers, these devices do not provide the contextual information needed to identify targets for behavioral interventions and generate public health guidelines to reduce sedentary behavior. Thus, self-reports still remain an important method of measurement for physical activity and sedentary behaviors.

Objective: This study evaluated the reliability, validity, and sensitivity to change of a smartphone app in assessing sitting, light-intensity physical activity (LPA), and moderate-vigorous physical activity (MVPA).
Methods: Adults (N=28; 49.0 years old, standard deviation [SD] 8.9; 85% men; 73% Caucasian; body mass index=35.0, SD 8.3 kg/m2) reported their sitting, LPA, and MVPA over an 11-week behavioral intervention. During three separate 7-day periods, participants wore the activPAL3c accelerometer/inclinometer as a criterion measure. Intraclass correlation (ICC; 95% CI) and bias estimates (mean difference [δ] and root of mean square error [RMSE]) were used to compare app-based reported behaviors to measured sitting time (lying/seated position), LPA (standing or stepping at <100 steps/minute), and MVPA (stepping at >100 steps/minute).

Results: Test-retest results suggested moderate agreement with the criterion for sedentary time, LPA, and MVPA (ICC=0.65 [0.43-0.82], 0.67 [0.44-0.83] and 0.69 [0.48-0.84], respectively). The agreement between the two measures was poor (ICC=0.05-0.40). The app underestimated sedentary time (δ=-45.9 [-67.6, -24.2] minutes/day, RMSE=201.6) and overestimated LPA and MVPA (δ=18.8 [-1.30 to 38.9] minutes/day, RMSE=183; and δ=29.3 [25.3 to 33.2] minutes/day, RMSE=71.6, respectively). The app underestimated change in time spent during LPA and MVPA but overestimated change in sedentary time. Both measures showed similar directions in changed scores on sedentary time and LPA.

Conclusions: Despite its inaccuracy, the app may be useful as a self-monitoring tool in the context of a behavioral intervention. Future research may help to clarify reasons for under- or over-reporting of behaviors.

There is an increasing awareness that health care must move from post-symptomatic treatment to presymptomatic intervention. An ideal system would allow regular inexpensive monitoring of health status using circulating antibodies to report on health fluctuations. Recently, we demonstrated that peptide microarrays can do this through antibody signatures (immunosignatures). Unfortunately, printed microarrays are not scalable. Here we demonstrate a platform based on fabricating microarrays (~10 M peptides per slide, 330,000 peptides per assay) on silicon wafers using equipment common to semiconductor manufacturing. The potential of these microarrays for comprehensive health monitoring is verified through the simultaneous detection and classification of six different infectious diseases and six different cancers. Besides diagnostics, these high-density peptide chips have numerous other applications both in health care and elsewhere.

Swinging arms are a key functional component of multistep catalytic transformations in many naturally occurring multi-enzyme complexes. This arm is typically a prosthetic chemical group that is covalently attached to the enzyme complex via a flexible linker, allowing the direct transfer of substrate molecules between multiple active sites within the complex. Mimicking this method of substrate channelling outside the cellular environment requires precise control over the spatial parameters of the individual components within the assembled complex. DNA nanostructures can be used to organize functional molecules with nanoscale precision and can also provide nanomechanical control. Until now, protein–DNA assemblies have been used to organize cascades of enzymatic reactions by controlling the relative distance and orientation of enzymatic components or by facilitating the interface between enzymes/cofactors and electrode surfaces. Here, we show that a DNA nanostructure can be used to create a multi-enzyme complex in which an artificial swinging arm facilitates hydride transfer between two coupled dehydrogenases. By exploiting the programmability of DNA nanostructures, key parameters including position, stoichiometry and inter-enzyme distance can be manipulated for optimal activity.