Experimental Evidence for the Role of Dynamics in pH-Dependent Enzymatic Activity.

Enzymatic activity is heavily influenced by pH, but the rationale for the dynamical mechanism of pH-dependent enzymatic activity has not been fully understood. In this work, combined neutron scattering techniques, including quasielastic neutron scattering (QENS) and small angle neutron scattering (SANS), are used to study the structural and dynamic changes of a model enzyme, xylanase, under different pH and temperature environments. The QENS results reveal that xylanase at optimal pH exhibits faster relaxational dynamics and a lower energy barrier between conformational substates. The SANS results demonstrate that pH affects both xylanase's stability and monodispersity. Our findings indicate that enzymes have optimized stability and function under their optimal pH conditions, with both structure and dynamics being affected. The current study offers valuable insights into enzymatic functionality mechanisms, allowing for broad industrial applications.

Prediction of available energy and amino acid digestibility of Chinese sorghum fed to growing-finishing pigs.

Two experiments were conducted to determine digestible energy (DE), metabolizable energy (ME), as well as the standardized ileal digestibility (SID) of crude protein (CP) and amino acids (AA) in 10 sorghum samples fed to pigs. In experiment 1, 22 crossbred barrows (Duroc × Yorkshire × Landrace, Initial body weight [BW]: 70.0 ±â€…1.8 kg) were selected and allotted to a replicated 11 × 3 incomplete Latin square design, including a basal diet and 10 sorghum energy diets and three consecutive periods. Each period had 7 d adaptation and 5 d total feces and urine collection. The DE and ME were determined by the total collection and the difference method. In experiment 2, 22 crossbred barrows (Duroc × Yorkshire × Landrace, Initial BW: 41.3 ±â€…1.2 kg) that had a T-cannula installed in the distal ileum were assigned to a replicated 11 × 3 incomplete Latin square design, including an N-free diet and 10 sorghum diets. Each period had 5 d adaptation and 2 d ileal digesta collection. The basal endogenous N losses were measured by the N-free diet method. All diets in experiment 2 were added 0.30% titanium dioxide as an indigestible marker for calculating the ileal CP and AA digestibility. On an as-fed basis, the DE and ME contents in sorghum were 3,410 kcal/kg (2,826 to 3,794 kcal/kg) and 3,379 kcal/kg (2,785 to 3,709 kcal/kg), respectively. The best-fit prediction equation for DE and ME were DE = 6,267.945 - (1,271.154 × % tannin) - (1,109.720 × % ash) (R2 = 0.803) and ME = 51.263 + (0.976 × DE) (R2 = 0.994), respectively. The SID of CP, Lys, Met, Thr, and Trp (SIDCP, SIDLys, SIDMet, SIDThr, and SIDTrp) in 10 sorghum samples were 78.48% (69.56% to 84.23%), 74.27% (61.11% to 90.60%), 92.07% (85.16% to 95.40%), 75.46% (66.39% to 80.80%) and 87.99% (84.21% to 92.37%), respectively. The best prediction equations for SID of CP and the first four limiting AAs were as following: SIDCP = 93.404 - (21.026 × % tannin) (R2 = 0.593), SIDCP = 42.922 - (4.011 × % EE) + (151.774 × % Met) (R2 = 0.696), SIDLys = 129.947 - (670.760 × % Trp) (R2 = 0.821), SIDMet = 111.347 - (232.298 × % Trp) (R2 = 0.647), SIDThr = 55.187 + (3.851 × % ADF) (R2 = 0.609) and SIDTrp = 95.676 - (10.824 × % tannin) (R2 = 0.523), respectively. Overall, tannin and ash are the first and second predictors of DE and ME values of sorghum, respectively, and the tannin, EE, Trp, ash, CF, and ADF can be used as the key predictors for SID of CP and first four limiting AAs.

In this manuscript, we selected and analyzed the chemical composition of 10 different varieties of Chinese sorghum. Two digestion and metabolism experiments were conducted to measure the concentrations of available energy (digestible energy and metabolizable energy) and ileal amino acid digestibility (apparent or standardized ileal amino acid digestibility) in sorghums fed to pigs. The results showed as following: 1) The available energy values of 10 sorghum samples varied greatly, and the ileal digestibility of some amino acids also differed. 2) The available energy value and the ileal digestibility of crude protein and amino acids can be predicted by the contents of chemical components in sorghum, such as tannin (a substance in sorghum that can bind to biological macromolecules like proteins), ash, tryptophan, and fiber composition, etc. The above findings enrich the ingredient database and assist for feed enterprises to make precise formulations when using the Chinese sorghum.

The Use of Hydrogel-Based Materials for Radioprotection.

Major causes of the radiation-induced disease include nuclear accidents, war-related nuclear explosions, and clinical radiotherapy. While certain radioprotective drug or bioactive compounds have been utilized to protect against radiation-induced damage in preclinical and clinical settings, these strategies are hampered by poor efficacy and limited utilization. Hydrogel-based materials are effective carriers capable of enhancing the bioavailability of compounds loaded therein. As they exhibit tunable performance and excellent biocompatibility, hydrogels represent promising tools for the design of novel radioprotective therapeutic strategies. This review provides an overview of common approaches to radioprotective hydrogel preparation, followed by a discussion of the pathogenesis of radiation-induced disease and the current states of research focused on using hydrogels to protect against these diseases. These findings ultimately provide a foundation for discussions of the challenges and future prospects associated with the use of radioprotective hydrogels.

Prognostic value of soluble urokinase-type plasminogen activator receptor in coronary artery disease: A meta-analysis.

BACKGROUND: A potential inflammatory biomarker, soluble urokinase-type plasminogen activator receptor (suPAR) has been utilized to assist the prognostic assessment of coronary artery disease (CAD) patients; however, outcomes have been inconsistent. The prognostic relevance of suPAR as a predictor of CAD patient adverse outcomes was therefore examined. METHODS: Research articles published as of 1 January 2022 were retrieved from PubMed, Embase, the Web of Science and the Cochrane Library. All-cause mortality, cardiovascular mortality and other major cardiovascular events (nonfatal myocardial infarction, heart failure or stroke) were analysed as a subset of relevant studies' results. We calculated hazard ratios (HRs) and 95% confidence intervals (CIs) for each study. The broad EQUATOR guidelines were conformed. Risk of bias was assessed with ROBINS-I tool. RESULTS: In total, this analysis included nine studies including 14,738 CAD patients. All included studies made a correction for certain potential confounders. However, risk of bias ranged from moderate to critical. When the ROBINS-I tool was used. Patients with CAD that exhibited increased suPAR levels had a substantially higher risk of all-cause mortality (HR = 2.24; 95% CI 1.97-2.55) or cardiovascular mortality (HR = 2.02; 95% CI 1.58-2.58), but not of developing other major cardiovascular events (HR = 1.63; 95% CI 0.86-3.11). Considerable heterogeneity across studies was observed in our meta-analyses, but no significant publication bias was detected. CONCLUSION: In patients with coronary disease, suPAR may have prognostic value for both all-cause and cardiovascular mortality but not for other major cardiovascular events.

Elastic Silicone Occlusive Sheeting Versus Silicone Occlusive Sheeting in the Treatment of Scars: A Randomized Controlled Trial.

INTRODUCTION: Hypertrophic scarring caused by conventional open thyroidectomy is prevalent among Asians and published trials have proved that silicone occlusive sheeting is a useful treatment for hypertrophic scarring. However, silicone occlusive sheeting does not effectively prevent scar widening. Here, we report elastic silicone occlusive sheeting as a new type of silicone application. In this study, we compared the effects of elastic silicone occlusive sheeting on scar width and appearance after conventional open thyroidectomy with those of silicone occlusive sheeting. METHODS: In this prospective, randomized, assessor-blinded study, a total of 74 patients who underwent conventional open thyroidectomy were recruited to undergo elastic silicone occlusive sheeting and silicone occlusive sheeting on the healed wound. Split scar study and scar quality were assessed on the basis of scar width, Vancouver scar scale, pain/itching visual analogue scale, and patients' subjective degree of satisfaction with the scar, during the patients' 6-month review. RESULTS: A total of 61 patients completed the study. Scar width, Vancouver scar scale score, and patients' subjective degree of satisfaction indicated that elastic silicone occlusive sheeting was associated with narrower scars and significant improvement in scar appearance. The two methods did not differ significantly with regard to pain/itching visual analogue scale. CONCLUSIONS: Our findings highlight elastic silicone occlusive sheeting as an effective treatment for scarring, resulting in narrower and better scars after conventional open thyroidectomy. The use of elastic silicone occlusive sheeting after conventional open thyroidectomy may minimize the formation of hypertrophic scars in the early postoperative period. TRIAL REGISTRATION: ChiCTR2100049740.

Design and Synthesis of a Mitochondria-Targeting Radioprotectant for Promoting Skin Wound Healing Combined with Ionizing Radiation Injury.

Wound healing is seriously retarded when combined with ionizing radiation injury, because radiation-induced excessive reactive oxygen species (ROS) profoundly affect cell growth and wound healing. Mitochondria play vital roles not only as cellular energy factories but also as the main source of endogenous ROS, and in this work a mitochondria-targeting radioprotectant (CY-TMP1) is reported for radiation injury-combined wound repair. It was designed, synthesized and screened out from different conjugates between mitochondria-targeting heptamethine cyanine dyes and a peroxidation inhibitor 2,2,6,6-tetramethylpiperidinyloxy (TEMPO). CY-TMP1 specifically accumulated in mitochondria, efficiently mitigated mitochondrial ROS and total intracellular ROS induced by 6 Gy of X-ray ionizing irradiation, thereby exhibiting a notable radioprotective effect. The mechanism study further demonstrated that CY-TMP1 protected mitochondria from radiation-induced injury, including maintaining mitochondrial membrane potential (MMP) and ATP generation, thereby reducing the ratio of cell apoptotic death. Particularly, an in vivo experiment showed that CY-TMP1 could effectively accelerate wound closure of mice after 6 Gy of whole-body ionizing radiation. Immunohistochemical staining further indicated that CY-TMP1 may improve wound repair through angiogenesis and re-epithelialization. Therefore, mitochondria-targeting ROS scavengers may present a feasible strategy to conquer refractory wound combined with radiation injury.

Investigations of structural and dynamical mechanisms of ice formation regulated by graphene oxide nanosheets.

Recent research indicates that graphene oxide (GO) nanosheets can be used to regulate ice formation by controlling critical ice nucleus growth in water at supercooling temperatures. In addition, the study of ice formation mechanisms regulated by GO nanosheets, a good model system for antifreeze proteins (AFPs), will shed light on how AFPs regulate ice formation in nature. In this work, time-resolved small-angle x-ray scattering (TR-SAXS) and quasi-elastic neutron scattering (QENS) experiments were carried out to investigate the structural and dynamical mechanisms of ice formation regulated by GO nanosheets. Strikingly, a transient intermediate state was observed in TR-SAXS experiments that only exists in the aqueous dispersions with a larger GO size (11 nm). This serves as evidence that the size of GO is critical for regulating ice formation. Elastic neutron scattering results indicate that ice is formed in all samples and thermal hysteresis occurs in GO aqueous dispersions in both H2O and D2O. The structural and dynamics information about water molecules in GO, extracted from QENS, reveals different dynamical behaviors of water molecules in GO aqueous dispersions when approaching the ice formation temperature.

Dynamic functional connectivity and its anatomical substrate reveal treatment outcome in first-episode drug-naïve schizophrenia.

Convergent evidence has suggested a significant effect of antipsychotic exposure on brain structure and function in patients with schizophrenia, yet the characteristics of favorable treatment outcome remains largely unknown. In this work, we aimed to examine how large-scale brain networks are modulated by antipsychotic treatment, and whether the longitudinal changes could track the improvements of psychopathologic scores. Thirty-four patients with first-episode drug-naïve schizophrenia and 28 matched healthy controls were recruited at baseline from Shanghai Mental Health Center. After 8 weeks of antipsychotic treatment, 24 patients were re-scanned. Through a systematical dynamic functional connectivity (dFC) analysis, we investigated the schizophrenia-related intrinsic alterations of dFC at baseline, followed by a longitudinal study to examine the influence of antipsychotic treatment on these abnormalities by comparing patients at baseline and follow-up. A structural connectivity (SC) association analysis was further carried out to investigate longitudinal anatomical changes that underpin the alterations of dFC. We found a significant symptomatic improvement-related increase in the occurrence of a dFC state characterized by stronger inter-network integration. Furthermore, symptom reduction was correlated with increased FC variability in a unique connectomic signature, particularly in the connections within the default mode network and between the auditory, cognitive control, and cerebellar network to other networks. Additionally, we observed that the SC between the superior frontal gyrus and medial prefrontal cortex was decreased after treatment, suggesting a relaxation of normal constraints on dFC. Taken together, these findings provide new evidence to extend the dysconnectivity hypothesis in schizophrenia from static to dynamic brain network. Moreover, our identified neuroimaging markers tied to the neurobiology of schizophrenia could be used as potential indicators in predicting the treatment outcome of antipsychotics.

Mesophilic Pyrophosphatase Function at High Temperature: A Molecular Dynamics Simulation Study.

The mesophilic inorganic pyrophosphatase from Escherichia coli (EcPPase) retains function at 353 K, the physiological temperature of hyperthermophilic Thermococcus thioreducens, whereas the homolog protein (TtPPase) from this hyperthermophilic organism cannot function at room temperature. To explain this asymmetric behavior, we examined structural and dynamical properties of the two proteins using molecular dynamics simulations. The global flexibility of TtPPase is significantly higher than its mesophilic homolog at all tested temperature/pressure conditions. However, at 353 K, EcPPase reduces its solvent-exposed surface area and increases subunit compaction while maintaining flexibility in its catalytic pocket. In contrast, TtPPase lacks this adaptability and has increased rigidity and reduced protein/water interactions in its catalytic pocket at room temperature, providing a plausible explanation for its inactivity near room temperature.

Frequency tuning ratio testing of a laser via a hollow photonic crystal fiber resonator.

In order to satisfy the requirements of laser frequency tuning ratio (FTR) measurement, experimental equipment based on a hollow photonic crystal fiber resonator (HPCFR) is proposed in this paper. First, the principle scheme of the equipment consisting of HPCFR is designed, and the resonance curves of the HPCFR are theoretically analyzed, calculated, and simulated; second, the transmissive HPCFR sample is fabricated and the resonance curve is obtained; eventually, the experimental results from the established laser FTR experimental setup demonstrate that the FTRs of a narrow-linewidth fiber laser and semiconductor laser are 17.6 MHz/V and 30.9 MHz/mA, respectively, which are basically in accordance with the factory parameters of the lasers. This work shows that the FTR experimental equipment via HPCFR has the advantages of high precision and good long-term stability.

Key Points of Preliminary Treatment for Patients with Extremity Trauma Based on Analysis of Amputation Cases.

INTRODUCTION: Severe trauma can lead to amputation of limbs. There is no golden standard or comprehensive evaluation indicator for amputation. It is difficult for the primary rescue organization to focus on the most essential indicators and to determine whether to perform amputation or take proper operation. PROBLEM: For medical staff in first-line medical teams for disaster relief or in a common primary hospital, what indicators should they focus on to keep the patient's limbs when they receive wounded patients with severe trauma? METHODS: A retrospective case-control study was performed based on the patients with severe trauma from January 2013 through December 2018 in the emergency department of Southwest Hospital (Shapingba District, Chongqing, China), a Level I trauma center. A total of 165 cases were divided into amputation group (n = 79) and non-amputation control group (n = 86), which had severe skin and muscle injury but without amputation. The causes of trauma and the special cases were analyzed. Binary logistic regression models were used to find the essential indicators for amputation. CONCLUSIONS: Neurovascular injury with delayed treatment was the most decisive indicator leading to amputation, and time phase was also important for limb salvage. Preliminary treatment of disaster victims and patients with severe trauma should focus on neurovascular status and timely delivery.

Functional recombinant single-chain variable fragment antibody against Agkistrodon acutus venom.

Agkistrodon acutus bites are conventionally treated with animal-derived antivenom, the use of which is limited due to allergic reactions and serum sickness. Thus in the present study, the genes of humanized antibodies produced in response to A. acutus venom were extracted from lymphocytes from patients bitten by A. acutus. A single-chain variable fragment (scFv) library against venom was constructed using a T7 phage display system. ScFv genes that exhibited high affinity to venom were selected by library biopanning. An expression system was constructed for antivenom scFv fused with 6×His tag at its N- and C-terminus using pET-28a (+) vector. The scFv proteins could achieve functional and soluble expression in Escherichia coli via the auto-induction method. The purity and activity of the scFv genes and proteins were confirmed by SDS-PAGE, western blotting and ELISA. The results indicated that three soluble scFv proteins exhibited specific affinity to A. acutus venom and were harvested via the auto-induction method.

MicroRNA-29b regulates hypertrophy of murine mesenchymal stem cells induced toward chondrogenesis.

OBJECTIVE: Chondrocyte hypertrophy, a terminal stage of chondrocyte differentiation, is essential to the endochondral bone formation and is one of the major pathological factors in osteoarthritis. This study investigated the role of microRNA-29b (miR-29b), which is involved in chondrogenesis, in the regulation of hypertrophy in chondrocytes. METHODS: miR-29b expression was assessed during murine mesenchymal stem cells (mMSCs) chondrogenesis. To detect whether miR-29b affects chondrocyte hypertrophy, the mMSCs induced toward chondrogenesis were transfected with miR-29b or its antisense inhibitor (antagomiR-29b). Finally, the differential effects of antagomiR-29b on chondrocytes at different differentiation stages were evaluated by loss-of-function experiments. RESULTS: miR-29b expression was low-level during the early chondrogenic differentiation, however, it was changed to high level during hypertrophy. Subsequently, the gain-of-function and loss-of-function experiments had confirmed that miR-29b promoted hypertrophy in mMSC-derived chondrocytes. In addition, we confirmed that on day 7, when cells were treated with antagomiR-29b, was the optimal intervention time for preventing hypertrophic phenotype of mMSCs in vitro. CONCLUSION: miR-29b regulated chondrogenesis homeostasis and enhance hypertrophic phenotype. These data suggest that miR-29b is a key regulator of the chondrocyte phenotype derived from mMSCs and it might be a potential target for articular cartilage repair.

Small-Angle Neutron Scattering Reveals Energy Landscape for Rhodopsin Photoactivation.

Knowledge of the activation principles for G-protein-coupled receptors (GPCRs) is critical to development of new pharmaceuticals. Rhodopsin is the archetype for the largest GPCR family, yet the changes in protein dynamics that trigger signaling are not fully understood. Here we show that rhodopsin can be investigated by small-angle neutron scattering (SANS) in fully protiated detergent micelles under contrast matching to resolve light-induced changes in the protein structure. In SANS studies of membrane proteins, the zwitterionic detergent [(cholamidopropyl)dimethylammonio]-propanesulfonate (CHAPS) is advantageous because of the low contrast difference between the hydrophobic core and hydrophilic head groups as compared with alkyl glycoside detergents. Combining SANS results with quasielastic neutron scattering reveals how changes in volumetric protein shape are coupled (slaved) to the aqueous solvent. Upon light exposure, rhodopsin is swollen by the penetration of water into the protein core, allowing interactions with effector proteins in the visual signaling mechanism.

Neutron scattering in the biological sciences: progress and prospects.

The scattering of neutrons can be used to provide information on the structure and dynamics of biological systems on multiple length and time scales. Pursuant to a National Science Foundation-funded workshop in February 2018, recent developments in this field are reviewed here, as well as future prospects that can be expected given recent advances in sources, instrumentation and computational power and methods. Crystallography, solution scattering, dynamics, membranes, labeling and imaging are examined. For the extraction of maximum information, the incorporation of judicious specific deuterium labeling, the integration of several types of experiment, and interpretation using high-performance computer simulation models are often found to be particularly powerful.

Cyanidin suppresses autophagic activity regulating chondrocyte hypertrophic differentiation.

Cartilage is a kind of special connective tissue which does not contain neither blood vessels nor lymphatics and nerves. Therefore, the damage in cartilage is difficult to be repaired spontaneously. Constructing tissue engineered cartilage provides a new technique for cartilage repairing. Mesenchymal stem cells (MSCs) possess a unique capability of self-renew and can differentiate into pre-chondrocytes which are frequently applied as seed cells in tissue engineering. However, in regenerated cartilage the chondrocytes derived from MSCs can hardly maintain homeostasis and preferentially present hypertrophic like phenotype. We investigated the effects of cyanidin, a natural organic compound, on chondrogenic and subsequent hypertrophic differentiation of MSCs in order to seek approaches to inhibit chondrocyte hypertrophy. We evaluated the effects of cyanidin on expression of chondrogenic and hypertrophic marker genes through RT-PCR, Western blot, alcian blue staining, and immunocytochemistry. The results showed that both chondrogenic related genes Sox9, Col2a1, and hypertrophic marker genes Runx2, Col10a1 were inhibited by cyanidin. In addition, we found that cyanidin promoted Nrf2 and p62 expression and suppressed LC3B expression during chondrogenic stage of MSCs. Meanwhile phosphorylation of IκBα and autophagosome related protein LC3B were inactivated by cyanidin during chondrocyte hypertrophic stage. Furthermore, rapamycin, an autophagy activator, abrogated the inhibitory effect of cyanidin on chondrogenic, and hypertrophic differentiation of MSCs. In conclusion, one potential mechanism of cyanidin, by which the chondrogenic and hypertrophic differentiation of MSCs were inhibited, was due to decreased autophagy activity. Our results indicated that cyanidin was a potential therapeutic agent for keeping mature chondrocyte functions.

Inhibitory effect of dihydroartemisinin on chondrogenic and hypertrophic differentiation of mesenchymal stem cells.

Chondrocytes located in hyaline cartilage may maintain phenotype while the chondrocytes situated in calcified cartilage differentiate into hypertrophy. Chondrogenic and hypertrophic differentiation of mesenchymal stem cells (MSCs) are two subsequent processes during endochondral ossification. However, it is necessary for chondrocytes to hold homeostasis and to inhibit hypertrophic differentiation in stem cell-based regenerated cartilage. Dihydroartemisinin (DHA) is derived from artemisia apiacea which has many biological functions such as anti-malarial and anti-tumor. Whereas the effects of DHA on chondrogenic and hypertrophic differentiation are poorly understand. In this study, the cytotoxicity of DHA was determined by CCK8 assay and the cell apoptosis was analyzed by flow cytometry. Additionally, the effects of DHA on chondrogenic and hypertrophic differentiation of MSCs are explored by RT-PCR, western blotting and immunohistochemistry. The results showed that DHA inhibited expression of chondrogenic markers including Sox9 and Col2a1 by activating Nrf2 and Notch signaling. After induced to chondrogenesis, cells were treated with hypertrophic induced medium with DHA. The results revealed that hypertrophic markers including Runx2 and Col10a1 were down-regulated following DHA treatment through Pax6/HOXA2 and Gli transcription factors. These findings indicate that DHA is negative to chondrogenesis and is protective against chondrocyte hypertrophy to improve chondrocytes stability. Therefore, DHA might be not suited for chondogenesis but be potential as a new therapeutic candidate to maintain the biological function of regenerated cartilage.

Hypertrophic differentiation of mesenchymal stem cells is suppressed by xanthotoxin via the p38­MAPK/HDAC4 pathway.

Chondrocyte hypertrophy is a physiological process in endochondral ossification. However, the hypertrophic­like alterations of chondrocytes at the articular surface may result in osteoarthritis (OA). In addition, the generation of fibrocartilage with a decreased biological function in tissue engineered cartilage, has been attributed to chondrocyte hypertrophy. Therefore, suppressing chondrocyte hypertrophy in OA and the associated regeneration of non­active cartilage is of primary concern. The present study examined the effects of xanthotoxin (XAT), which is classified as a furanocoumarin, on chondrocyte hypertrophic differentiation of mesenchymal stem cells. Following XAT treatment, the expression levels of genes associated with chondrocyte hypertrophy were detected via immunohistochemistry, western blotting and reverse transcription­quantitative polymerase chain reaction. The results revealed that XAT inhibited the expression of various chondrocyte hypertrophic markers, including runt related transcription factor 2 (Runx2), matrix metalloproteinase 13 and collagen type X α1 chain. Further exploration indicated that XAT reduced the activation of p38­mitogen activated protein kinase and then increased the expression of histone deacetylase 4 to suppress Runx2. The findings indicated that XAT maintained the chondrocyte phenotype in regenerated cartilage and therefore may exhibit promise as a potential drug for the treatment of OA in the future.

Collective Excitations in Protein as a Measure of Balance Between its Softness and Rigidity.

In this article, we elucidate the protein activity from the perspective of protein softness and flexibility by studying the collective phonon-like excitations in a globular protein, human serum albumin (HSA), and taking advantage of the state-of-the-art inelastic X-ray scattering (IXS) technique. Such excitations demonstrate that the protein becomes softer upon thermal denaturation due to disruption of weak noncovalent bonds. On the other hand, no significant change in the local excitations is detected in ligand- (drugs) bound HSA compared to the ligand-free HSA. Our results clearly suggest that the protein conformational flexibility and rigidity are balanced by the native protein structure for biological activity.

Quasi-elastic Neutron Scattering Reveals Ligand-Induced Protein Dynamics of a G-Protein-Coupled Receptor.

Light activation of the visual G-protein-coupled receptor (GPCR) rhodopsin leads to significant structural fluctuations of the protein embedded within the membrane yielding the activation of cognate G-protein (transducin), which initiates biological signaling. Here, we report a quasi-elastic neutron scattering study of the activation of rhodopsin as a GPCR prototype. Our results reveal a broadly distributed relaxation of hydrogen atom dynamics of rhodopsin on a picosecond-nanosecond time scale, crucial for protein function, as only observed for globular proteins previously. Interestingly, the results suggest significant differences in the intrinsic protein dynamics of the dark-state rhodopsin versus the ligand-free apoprotein, opsin. These differences can be attributed to the influence of the covalently bound retinal ligand. Furthermore, an idea of the generic free-energy landscape is used to explain the GPCR dynamics of ligand-binding and ligand-free protein conformations, which can be further applied to other GPCR systems.