A Slow Conformational Switch in the BMAL1 Transactivation Domain Modulates Circadian Rhythms.

The C-terminal transactivation domain (TAD) of BMAL1 (brain and muscle ARNT-like 1) is a regulatory hub for transcriptional coactivators and repressors that compete for binding and, consequently, contributes to period determination of the mammalian circadian clock. Here, we report the discovery of two distinct conformational states that slowly exchange within the dynamic TAD to control timing. This binary switch results from cis/trans isomerization about a highly conserved Trp-Pro imide bond in a region of the TAD that is required for normal circadian timekeeping. Both cis and trans isomers interact with transcriptional regulators, suggesting that isomerization could serve a role in assembling regulatory complexes in vivo. Toward this end, we show that locking the switch into the trans isomer leads to shortened circadian periods. Furthermore, isomerization is regulated by the cyclophilin family of peptidyl-prolyl isomerases, highlighting the potential for regulation of BMAL1 protein dynamics in period determination.

The Bright Ideas TBI Camp: fostering innovation in interprofessional education and collaborative practice for traumatic brain injury by students in rehabilitation professions.

This report describes an innovative interprofessional education collaborative practice (IPCP) experience for rehabilitation professions students using a unique on-campus camp model through a community-academic partnership. Throughout the three-day camp, known as the Bright Ideas TBI Camp, interprofessional student groups deliver tailored health and wellness services to individuals with disabilities due to traumatic brain injury and their caregivers. Initial program evaluation suggests that this camp model offers an effective IPCP experience for students while addressing community health needs. Further outcome evaluation is needed to determine the impact of the camp on students' development of IPCP competencies and health outcomes of clients and caregivers.

Genetic variability of oxidative stress and DNA repair genes associated with pre-treatment cancer-related fatigue in women with breast cancer.

PURPOSE: Investigate potential relationships between pre-treatment cancer-related fatigue (CRF) in women with early-stage breast cancer and variation in genes involved with oxidative stress and DNA repair. METHODS: Investigated 39 functional and tagging single nucleotide polymorphisms (SNPs) in genes involved in oxidative stress (CAT, GPX1, SEPP1, SOD1, and SOD2) and DNA repair (ERCC2, ERCC3, ERCC5, and PARP1) in a sample (N = 219) that included n = 138 postmenopausal women diagnosed with early-stage breast cancer before initiation of therapy and n = 81 age- and education-matched healthy controls. Using the Profile of Mood States Fatigue/Inertia Subscale, fatigue occurrence and severity were evaluated in both groups. Regression analysis was used to independently identify significant SNPs for three outcomes: 1) any fatigue versus no fatigue, 2) clinically meaningful versus non-clinically meaningful fatigue, and 3) fatigue severity. Using a weighted multi-SNP method, genetic risk scores (GRS) were calculated for each participant, and GRS models were constructed for each outcome. Models were adjusted for age, pain, and symptoms of depression and anxiety. RESULTS: SEPP1rs3877899, ERCC2rs238406, ERCC2rs238416, ERCC2rs3916874, and ERCC3rs2134794 were associated with fatigue occurrence and had a significant GRS model (OR = 1.317, 95%CI [1.067, 1.675], P ≤ 0.05). One SNP, SOD2rs5746136, was significant for clinically meaningful fatigue; therefore, a GRS model could not be constructed. ERCC3rs4150407, ERCC3rs4150477, and ERCC3rs2134794 were associated with fatigue severity with a significant GRS model (b = 1.010, 95%CI [1.647, 4.577], R2 = 6.9%, P ≤ 0.01). CONCLUSIONS: These results may contribute to identifying patients who are at risk of developing CRF. Oxidative stress and DNA repair biological pathways may be involved with CRF.

Electrophysiological Measures of Swallowing Functions: A Systematic Review.

The purpose of this systematic review was to examine the application of event-related potentials (ERPs) to investigate neural processes of swallowing functions in adults with and without dysphagia. Computerized literature searches were performed from three search engines. Studies were screened using Covidence (Cochrane tool) and followed the Preferred Reporting Items for Systematic Reviews and Meta-Analyses statement standards (PRISMA-2009). A total of 759 studies were initially retrieved, of which 12 studies met inclusion criteria. Electrophysiological measures assessing swallowing functions were identified in two major ERP categories: (1) sensory potentials and (2) pre-motor potentials. Approximately 80% of eligible studies demonstrated strong methodological quality, although most employed a case series or case-control study design. Pharyngeal sensory-evoked potentials (PSEPs) were used to assess pharyngeal afferent cortical processing. The temporal sequence of the PSEP waveforms varied based on the sensory stimuli. PSEPs were delayed with localized scalp maps in patients with dysphagia as compared to healthy controls. The pre-motor ERPs assessed the cortical substrates involved in motor planning for swallowing, with the following major neural substrates identified: pre-motor cortex, supplementary motor area, and primary sensorimotor cortex. The pre-motor ERPs differed in amplitude for the swallow task (saliva versus liquid swallow), and the neural networks differed for cued versus non-cued task of swallowing suggesting differences in cognitive processes. This systematic review describes the application of electrophysiological measures to assess swallowing function and the promising application for furthering understanding of the neural substrates of swallowing. Standardization of protocols for use of electrophysiological measures to examine swallowing would allow for aggregation of study data to inform clinical practice for dysphagia rehabilitation.

The spliceosomal proteins PPIH and PRPF4 exhibit bi-partite binding.

Pre-mRNA splicing is a dynamic, multistep process that is catalyzed by the RNA (ribonucleic acid)-protein complex called the spliceosome. The spliceosome contains a core set of RNAs and proteins that are conserved in all organisms that perform splicing. In higher organisms, peptidyl-prolyl isomerase H (PPIH) directly interacts with the core protein pre-mRNA processing factor 4 (PRPF4) and both integrate into the pre-catalytic spliceosome as part of the tri-snRNP (small nuclear RNA-protein complex) subcomplex. As a first step to understand the protein interactions that dictate PPIH and PRPF4 function, we expressed and purified soluble forms of each protein and formed a complex between them. We found two sites of interaction between PPIH and the N-terminus of PRPF4, an unexpected result. The N-terminus of PRPF4 is an intrinsically disordered region and does not adopt secondary structure in the presence of PPIH. In the absence of an atomic resolution structure, we used mutational analysis to identify point mutations that uncouple these two binding sites and find that mutations in both sites are necessary to break up the complex. A discussion of how this bipartite interaction between PPIH and PRPF4 may modulate spliceosomal function is included.

P300 Event-Related Potentials Differentiate Better Performing Individuals With Traumatic Brain Injury: A Preliminary Study of Semantic Processing.

OBJECTIVE: To measure the effect of traumatic brain injury on the cognitive processing of words, as measured by the P300, in a semantic categorization task. PARTICIPANTS: Eight adults with a history of moderate to severe traumatic brain injury and 8 age- and gender-matched controls. DESIGN: A pilot study measuring cognitive event-related potentials in response to word pairs that were either in same or different semantic categories. MAIN MEASURES: The P300 (P3b) component of the auditory event-related potential and neuropsychological assessment. RESULTS: Two patterns of P300 amplitude related to brain injury were observed. Participants with poorer performance on neuropsychological tests exhibited reduced P300 amplitude as compared to controls but showed the typical P300 parietal scalp distribution. In contrast, better performing participants demonstrated robust P300 amplitude but a substantially altered scalp distribution, characterized by the recruitment of anterior brain regions in addition to parietal activation. CONCLUSIONS: The recruitment of frontal areas after traumatic brain injury may represent compensatory neural mechanisms utilized to successfully maximize task performance. The P300 in a semantic processing paradigm may be a sensitive marker of neural plasticity that could be used to improve functional outcomes in cognitive remediation paradigms.

Recruitment Strategies and Costs Associated With Enrolling People With Insomnia and High Blood Pressure Into an Online Behavioral Sleep Intervention: A Single-Site Pilot Study.

BACKGROUND: Recruitment in clinical research is a common challenge and source of study failure. The reporting of recruitment methods and costs in hypertension trials is limited especially for smaller, single-site trials, online intervention trials, and trials using newer online recruitment strategies. OBJECTIVE: The aims of this study are to describe and examine the feasibility of newer online-e-mail recruitment strategies and traditional recruitment strategies used to enroll participants with insomnia and high blood pressure into an online behavioral sleep intervention study (Sleeping for Heart Health). METHODS: The 16 online-e-mail-based and traditional recruitment strategies used are described. Recruitment strategy feasibility was examined by study interest and enrollee yields, conversion rates, and costs (direct, remuneration, labor, and cost per enrollee). RESULTS: From August 2014 to October 2015, 183 people were screened and 58 (31.7%) enrolled in the study (51.1 ± 12.9 years, 63.8% female, 72.4% African American, 136 ± 12/88 ± 7 mm Hg, 87.9% self-reported hypertension, 67.2% self-reported antihypertensive medication use). The recruitment strategies yielding the highest enrollees were the university hospital phone waiting message system (25.4%), Craigslist (22.4%), and flyers (20.3%) at a per enrollee cost of $42.84, $98.90, and $128.27, respectively. The university hospital phone waiting message system (55.6%) and flyers (54.5%) had the highest interested participant to enrolled participant conversion rate of all recruitment strategies. CONCLUSION: Approximately 70% of all enrolled participants were recruited from the university hospital phone waiting message system, Craigslist, or flyers. Given the recruitment challenges that most researchers face, we encourage the documenting, assessing, and reporting of detailed recruitment strategies and associated recruitment costs so that other researchers may benefit.

Nuclear cyclophilins affect spliceosome assembly and function in vitro.

Cyclophilins are ubiquitously expressed proteins that bind to prolines and can catalyse cis/trans isomerization of proline residues. There are 17 annotated members of the cyclophilin family in humans, ubiquitously expressed and localized variously to the cytoplasm, nucleus or mitochondria. Surprisingly, all eight of the nuclear localized cyclophilins are found associated with spliceosomal complexes. However, their particular functions within this context are unknown. We have therefore adapted three established assays for in vitro pre-mRNA splicing to probe the functional roles of nuclear cyclophilins in the context of the human spliceosome. We find that four of the eight spliceosom-associated cyclophilins exert strong effects on splicing in vitro. These effects are dose-dependent and, remarkably, uniquely characteristic of each cyclophilin. Using both qualitative and quantitative means, we show that at least half of the nuclear cyclophilins can act as regulatory factors of spliceosome function in vitro. The present work provides the first quantifiable evidence that nuclear cyclophilins are splicing factors and provides a novel approach for future work into small molecule-based modulation of pre-mRNA splicing.

Age-Related Differences in the Effects of Linguistic and Nonlinguistic Masking on Semantic Processing: Evidence From the N400 Component in Young and Middle-Aged Listeners.

PURPOSE: The purpose of this study was to determine if there are age-related differences in semantic processing with linguistic and nonlinguistic masking, as measured by the N400. METHOD: Sixteen young (19-31 years) and 16 middle-aged (41-57 years) adults with relatively normal hearing sensitivity were asked to determine whether word pairs were semantically related or unrelated in three listening conditions: quiet, forward, and reverse two-talker speech competition at 0 dB SNR. Behavioral data (accuracies and reaction times) and auditory event-related potential data (N400 amplitudes and latencies) were analyzed using separate mixed design multivariate analysis of variances. RESULTS: Mean N400 amplitudes for semantically related word pairs were similar between young and middle-aged adults. Although neither group showed N400 amplitude differences between masker types, N400 amplitude was significantly greater in the presence of linguistic and nonlinguistic masking than in quiet. In contrast, mean N400 amplitudes for semantically unrelated words were significantly more negative for young adults and not significantly different among listening conditions. CONCLUSIONS: Our findings illustrated age-related differences during a semantic processing task, as indexed by the N400, that may not be evident in suprathreshold speech repetition/recognition tasks or behavioral data. Additionally, N400 amplitudes indicated that linguistic masking effects were equivalent to nonlinguistic masking effects on semantic processing.

Association Between Genes in the Nuclear Factor E2-Related Factor 2 Antioxidative Response Elements Pathway and Cancer-Related Fatigue in Women With Early-Stage Breast Cancer.

OBJECTIVES: To explore genes in the nuclear factor E2-related factor 2 antioxidative response elements (Nrf2-ARE) signaling pathway using a multiomics approach for associations with variability of cancer-related fatigue (CRF) in postmenopausal women with early-stage hormone receptor-positive breast cancer. SAMPLE & SETTING: Postmenopausal women (N = 116) with early-stage hormone receptor-positive breast cancer were recruited from western Pennsylvania. METHODS & VARIABLES: Candidate genes from the Nrf2-ARE pathway were investigated for associations with CRF occurrence and severity. Associations were evaluated using logistic regression for occurrence and linear regression for severity. RESULTS: The rs2706110 TT genotype in NFE2L2 was associated with a 3.5-fold increase in odds of CRF occurrence. The cytosine-phosphate-guanine (CpG) site cg22820568 in PRDX1 was associated with CRF occurrence and severity. IMPLICATIONS FOR NURSING: Biomarkers based on Nrf2-ARE genes may help to identify women at increased risk for more severe CRF and to develop targeted interventions.

Structural and biochemical characterization of the human cyclophilin family of peptidyl-prolyl isomerases.

Peptidyl-prolyl isomerases catalyze the conversion between cis and trans isomers of proline. The cyclophilin family of peptidyl-prolyl isomerases is well known for being the target of the immunosuppressive drug cyclosporin, used to combat organ transplant rejection. There is great interest in both the substrate specificity of these enzymes and the design of isoform-selective ligands for them. However, the dearth of available data for individual family members inhibits attempts to design drug specificity; additionally, in order to define physiological functions for the cyclophilins, definitive isoform characterization is required. In the current study, enzymatic activity was assayed for 15 of the 17 human cyclophilin isomerase domains, and binding to the cyclosporin scaffold was tested. In order to rationalize the observed isoform diversity, the high-resolution crystallographic structures of seven cyclophilin domains were determined. These models, combined with seven previously solved cyclophilin isoforms, provide the basis for a family-wide structure:function analysis. Detailed structural analysis of the human cyclophilin isomerase explains why cyclophilin activity against short peptides is correlated with an ability to ligate cyclosporin and why certain isoforms are not competent for either activity. In addition, we find that regions of the isomerase domain outside the proline-binding surface impart isoform specificity for both in vivo substrates and drug design. We hypothesize that there is a well-defined molecular surface corresponding to the substrate-binding S2 position that is a site of diversity in the cyclophilin family. Computational simulations of substrate binding in this region support our observations. Our data indicate that unique isoform determinants exist that may be exploited for development of selective ligands and suggest that the currently available small-molecule and peptide-based ligands for this class of enzyme are insufficient for isoform specificity.

Electrophysiological evidence of augmented interaural asymmetry in middle-aged listeners.

BACKGROUND: Various dimensions of auditory processing, especially the perception of speech in the presence of background competition, have been shown to deteriorate with age. A persistent problem in the assessment of these age-related changes has been the high prevalence of age-related high-frequency hearing loss in elderly persons. Some investigators have suggested that a more fruitful approach to the study of age-related decline might be to study middle-aged, rather than elderly, persons, where confounding high-frequency hearing loss is less prevalent. PURPOSE: To determine whether an increase in the left-ear disadvantage (LED) in dichotic listening could be demonstrated in a group of middle-aged persons. RESEARCH DESIGN: The N400 component of the auditory event-related potential (AERP) was utilized to evaluate interaural asymmetry in a quasi-dichotic competing speech task. Electrophysiological responses were obtained on a word-pair semantic categorization task presented through a front loudspeaker while the listener ignored competing speech presented through either left (competition left [CL]) or right (competition right [CR]) loudspeakers. Study Samples: Twenty young (18-24 yr) and 20 middle-aged (44-57 yr) females with normal hearing sensitivity. DATA COLLECTION AND ANALYSIS: Individual, as well as grand-averaged, AERP waveforms and scalp topographies were analyzed for the word pairs. Peak amplitude and latency measures of the N400 component were subjected to a mixed design analysis of variance (ANOVA). RESULTS: No significant interaural asymmetry was found in the AERP waveform for the reference word condition in either age group. In response to the second word of the pair, however, middle-aged females showed significantly greater N400 negativity in the CR condition than in the CL condition. No significant laterality effect was found in the young females. CONCLUSIONS: The study of young versus middle-aged participants may be an effective way of avoiding the confound of high-frequency hearing loss in elderly persons when studying age effects on auditory processing.

Alteration of the Chicken Upper Respiratory Microbiota, Following H9N2 Avian Influenza Virus Infection.

Several studies have highlighted the importance of the gut microbiota in developing immunity against viral infections in chickens. We have previously shown that H9N2 avian influenza A virus (AIV) infection retards the diversity of the natural colon-associated microbiota, which may further influence chicken health following recovery from infection. The effects of influenza infection on the upper respiratory tract (URT) microbiota are largely unknown. Here, we showed that H9N2 AIV infection lowers alpha diversity indices in the acute phase of infection in the URT, largely due to the family Lactobacillaceae being highly enriched during this time in the respiratory microbiota. Interestingly, microbiota diversity did not return to levels similar to control chickens in the recovery phase after viral shedding had ceased. Beta diversity followed a similar trend following the challenge. Lactobacillus associate statistically with the disturbed microbiota of infected chickens at the acute and recovery phases of infection. Additionally, we studied age-related changes in the respiratory microbiota during maturation in chickens. From 7 to 28 days of age, species richness and evenness were observed to advance over time as the microbial composition evolved. Maintaining microbiota homeostasis might be considered as a potential therapeutic target to prevent or aid recovery from H9N2 AIV infection.

Epigenomic Links Between Social Determinants of Health and Symptoms: A Scoping Review.

Social determinants of health (SDoH) impact health and wellness. The link between SDoH and adverse health outcomes, including symptom occurrence and severity, may be explained by an individual's physiologic response to one or more SDoH. One potential mechanism underlying this physiologic response linking SDoH and symptoms is the dynamic epigenome. The purpose of this scoping review of the literature was to examine differential susceptibility for symptoms by identifying and summarizing research linking SDoH and symptoms through epigenomic mechanisms. PubMed was searched to identify empirical research where at least one SDoH was an independent or dependent variable, at least one symptom was investigated, and the investigation included an epigenomic measure. Of the 484 articles initially retrieved, after thorough vetting, 41 articles met eligibility. The most studied symptom was depressive symptoms followed by anxiety, cognitive function, sleep dysfunction, and pain. The most frequently studied SDoH were: 1) stress, particularly early life stress and acculturative stress; and 2) trauma, predominantly childhood trauma. DNA methylation and telomere length were the most studied epigenomic measures. Four genes (SLC6A4, BDNF, NR3C1, OXTR) had evidence from multiple studies and across methodological approaches linking SDoH to symptoms. This review supports the inclusion of epigenomic approaches to better understand the link between SDoH and symptoms and provides evidence that SDoH impact telomere length and the methylation of genes involved in neurotransmitter signaling, neuronal survival, behavior, inflammation and stress response.

Brain-derived neurotrophic factor (BDNF) epigenomic modifications and brain-related phenotypes in humans: A systematic review.

Epigenomic modifications of the brain-derived neurotrophic factor (BDNF) gene have been postulated to underlie the pathogenesis of neurodevelopmental, psychiatric, and neurological conditions. This systematic review summarizes current evidence investigating the association of BDNF epigenomic modifications (DNA methylation, non-coding RNA, histone modifications) with brain-related phenotypes in humans. A novel contribution is our creation of an open access web-based application, the BDNF DNA Methylation Map, to interactively visualize specific positions of CpG sites investigated across all studies for which relevant data were available. Our literature search of four databases through September 27, 2021 returned 1701 articles, of which 153 met inclusion criteria. Our review revealed exceptional heterogeneity in methodological approaches, hindering the identification of clear patterns of robust and/or replicated results. We summarize key findings and provide recommendations for future epigenomic research. The existing literature appears to remain in its infancy and requires additional rigorous research to fulfill its potential to explain BDNF-linked risk for brain-related conditions and improve our understanding of the molecular mechanisms underlying their pathogenesis.

Auditory-cognitive interactions underlying interaural asymmetry in an adult listener: a case study.

OBJECTIVE: Abnormal interaural asymmetry on tests of dichotic listening is commonly observed in individuals suspected of auditory processing disorder (APD). Although a structural basis for the abnormality has been widely accepted, the influence of cognitive variables on the degree of observed asymmetry has gained increasing attention. To study this issue, we manipulated cognitive influences on interaural asymmetry in an adult with the auditory complaints typically associated with APD. STUDY SAMPLE: A 55 year-old woman with complaints of difficulty understanding speech in noisy environments despite normal audiometric levels. DESIGN: Several experimental dichotic procedures were administered. Each procedure was characterized by the manipulation of cognitive task demands. RESULTS: Interaural asymmetry was greatest when the demands on attention and/or memory were maximal. Electrophysiological data revealed interaural asymmetry on later stages of information processing. CONCLUSIONS: Results are discussed in relation to auditory-specific outcomes on clinical tests for APD.

Interprofessional Collaborative Practice Trends Between Speech-Language Pathologists and Audiologists.

PURPOSE: This study investigated interprofessional practice (IPP) trends and attitudes between speech-language pathologists (SLP) and audiologists (AUD). METHODS: Clinical SLPs and AUDs completed an online survey, consisting of demographics, caseload trends, collaborative practice trends, and barriers to collaborative practice. RESULTS: A total of 237 participants (131 SLPs, 106 AUDs) completed an online survey. Survey results indicated that IPP is important or very important to both professions. A smaller proportion of SLPs reported collaborating with AUDs than AUDs reported collaborating with SLPs. The most frequently identified barrier to collaboration experienced by the two professions was "access." Based on their beliefs, however, the top barriers differed between the professions. Specifically, SLPs believed that "access" was the top barrier to collaboration with AUDs whereas AUDs believed that "attitudes and perceptions" was the top barrier to collaboration with SLPs. Findings from the revised Attitudes Towards Health Care Teams Scale indicated that SLPs had a significantly more positive attitude toward IPP than AUDs. CONCLUSIONS: This study indicated that although IPP is valued by SLPs and AUDs, barriers continue to limit the percentage of practitioners engaging in IPP. Although both professions experienced common barriers, their beliefs about barriers to collaboration differed.

A Preliminary Exploration of Pitch Discrimination, Temporal Sequencing, and Prosodic Awareness Skills of Children Who Participate in Different School-Based Music Curricula.

Musical training has been shown to have a positive influence on a variety of skills, including auditory-based tasks and nonmusical cognitive and executive functioning tasks; however, because previous investigations have yielded mixed results regarding the relationship between musical training and these skills, the purpose of this study was to examine and compare the auditory processing skills of children who receive focused, daily musical training with those with more limited, generalized musical training. Sixteen typically developing children (second-fourth grade) from two different schools receiving different music curricula were assessed on measures of pitch discrimination, temporal sequencing, and prosodic awareness. The results indicated significantly better scores in pitch discrimination abilities for the children receiving daily, focused musical training (School 1) compared to students attending music class only once per week, utilizing a more generalized elementary school music curriculum (School 2). The findings suggest that more in-depth and frequent musical training may be associated with better pitch discrimination abilities in children. This finding is important given that the ability to discriminate pitch has been linked to improved phonological processing skills, an important skill for developing spoken language and literacy. Future investigations are needed to determine whether the null findings for temporal sequencing and prosodic awareness can be replicated or may be different for various grades and tasks for measuring these abilities.

A conserved mechanism of autoinhibition for the AMPK kinase domain: ATP-binding site and catalytic loop refolding as a means of regulation.

The AMP-activated protein kinase (AMPK) is a highly conserved trimeric protein complex that is responsible for energy homeostasis in eukaryotic cells. Here, a 1.9 A resolution crystal structure of the isolated kinase domain from the alpha2 subunit of human AMPK, the first from a multicellular organism, is presented. This human form adopts a catalytically inactive state with distorted ATP-binding and substrate-binding sites. The ATP site is affected by changes in the base of the activation loop, which has moved into an inhibited DFG-out conformation. The substrate-binding site is disturbed by changes within the AMPKalpha2 catalytic loop that further distort the enzyme from a catalytically active form. Similar structural rearrangements have been observed in a yeast AMPK homologue in response to the binding of its auto-inhibitory domain; restructuring of the kinase catalytic loop is therefore a conserved feature of the AMPK protein family and is likely to represent an inhibitory mechanism that is utilized during function.

Autoregulation by the juxtamembrane region of the human ephrin receptor tyrosine kinase A3 (EphA3).

Ephrin receptors (Eph) affect cell shape and movement, unlike other receptor tyrosine kinases that directly affect proliferative pathways. The kinase domain of EphA3 is activated by ephrin binding and receptor oligomerization. This activation is associated with two tyrosines in the juxtamembrane region; these tyrosines are sites of autophosphorylation and interact with the active site of the kinase to modulate activity. This allosteric event has important implications both in terms of understanding signal transduction pathways mediated by Eph kinases as well as discovering specific therapeutic ligands for receptor kinases. In order to provide further details of the molecular mechanism through which the unphosphorylated juxtamembrane region blocks catalysis, we studied wild-type and site-specific mutants in detail. High-resolution structures of multiple states of EphA3 kinase with and without the juxtamembrane segment allowed us to map the coupled pathway of residues that connect the juxtamembrane segment, the activation loop, and the catalytic residues of the kinase domain. This highly conserved set of residues likely delineates a molecular recognition pathway for most of the Eph RTKs, helping to characterize the dynamic nature of these physiologically important enzymes.