Sensitivity improvement of a single-NV diamond magnetometer using a chiral waveguide.

A single nitrogen-vacancy (NV) center in diamond is utilized to perform nanoscale magnetic measurements. However, the low contrast and poor collection efficiency of spin-dependent emitted photons limited the instrument sensitivity to approximately several nT/Hz. Here, we design a diamond magnetometer based on a chiral waveguide. We numerically demonstrate that the proposed device achieves a sensitivity of 170 pT/Hz owing to near-unity contrast and efficient photon collection. We also confirm that the device sensitivity is robust against position misalignment and dipole misorientation of an NV center. The proposed approach will enable the construction of a highly-sensitive magnetometer with high spatial resolution.

The morphology of the femoral posterior condyle affects the external rotation of the femur.

PURPOSE: The purpose of this study was to identify factors related to the external rotation of the femur during knee flexion. METHODS: Three-dimensional (3D) digital models of the femur and tibia were reconstructed from computed tomography images of 41 healthy Japanese subjects. Thirteen parameters related to femoral and tibial morphology and alignment of the lower extremities were evaluated, including the inclination angle of the posterior lateral and medial femoral condyles, the ratio of the medial and lateral posterior condyle radii approximated as spheres, the spherical condylar angle, the posterior condylar angle, the medial and lateral posterior tibial slope, the difference of medial and lateral posterior tibial slope, the tibiofemoral rotation angle, the 3D femorotibial angle, the 3D hip-knee-ankle angle, and the passing point of the weight-bearing line (medial-lateral and anterior-posterior). The rotation angle of the femur relative to the tibia during squatting was investigated using a 3D to 2D image matching technique and the relationships with the13 parameters were determined. RESULTS: The femur externally rotated substantially up to 20° of knee flexion (9.2° ± 3.7°) and gently rotated after 20° of knee flexion (12.8° ± 6.2°). The external rotation angle at 20°-120° of knee flexion correlated with the spherical condylar angle, the tibiofemoral rotation angle and the inclination angle of the posterior medial condyles (correlation coefficient; 0.506, 0.364, 0.337, respectively). CONCLUSION: The parameter that was most related to the external rotation of the femur during knee flexion was the spherical condylar angle. LEVEL OF EVIDENCE: IV.

Variations in the Photoexcitation Mechanism of an Adsorbed Molecule on a Gold Nanocluster Governed by Interfacial Contact.

We performed first-principles calculations on the optical response of a Au147-azobenzene complex to elucidate the role of the interfacial contact between Au147 and azobenzene. Our calculations of photoexcited electron dynamics for different configurations of azobenzene adsorbed on the Au147 nanocluster revealed that the optical properties of the azobenzene moiety change markedly by the interfacial contact, even if the electronic structure in the ground state is almost unchanged. The optical absorption measured for isolated azobenzene weakens when the Au147-azobenzene interaction increases, while the absorption measured using the light field along the Au147-azobenzene alignment strengthens. The electronic excitation analysis showed that the mechanism of the charge-transfer excitation between Au147 and azobenzene changes remarkably depending on the strength of the interfacial interaction. We revealed that the optical property can be governed by the atomic-scale difference in the adsorption structure of azobenzene on a Au147 nanocluster. This study affords novel insights that could enable the photoexcitation mechanism to be controlled by designing the interface between a metal nanoparticle and a molecule.

The Influence of Knee Osteoarthritis on Spinopelvic Alignment and Global Sagittal Balance.

Osteoarthritis (OA) of the knee is thought to lead to a loss of lumbar lordosis (LL) as a compensation for knee flexion contracture. Changes in sagittal alignment are not limited to the lumbar spine and involve a complex interplay of alignment of the hip, pelvis, and spine. While spine-hip interactions have been previously explored, the influence of knee OA sagittal alignment parameters on spinopelvic alignment and global sagittal balance remains unexplored. Standing radiological examination using EOS biplanar radiography was examined in 108 patients with knee OA. Whole-body sagittal alignment parameters (thoracic kyphosis, LL, pelvic incidence, pelvic tilt [PT], femoropelvic angle [FPA], femoral tilt angle [FTA], tibial tilt angle, and knee flexion angle [KFA]) and global balance parameters (sagittal vertical axis [SVA] and odontoid hip axis [OD-HA] angle) were measured three dimensionally (3D). The correlation coefficients among all parameters were assessed. A multiple stepwise linear regression model was built to investigate the direct association between SVA or OD-HA angle (dependent variables) and sagittal alignment parameters and demographic data (independent variables). Significant correlations between KFA, FPA, FTA, SVA, and OD-HA angle were found. FTA was correlated with LL and FPA. The FTA was the most influential predictor of both global sagittal balance parameters (p < 0.001). Knee OA leads to changes in global sagittal balance with effects at the hip, knee, pelvis, and spine. FTA (forward flexion of the femur vs. the vertical plane) is the largest driver of global sagittal plane balance in patients with knee OA.

Postoperative femoral anteroposterior position while standing correlates with the posterior tibial slope and posterior femoral condylar offset in medial pivot total knee arthroplasty.

BACKGROUND: After total knee arthroplasty (TKA), the femur tends to be located posteriorly under weight-bearing (WB) conditions, and a paradoxical femoral anterior motion occurs, leading to joint instability. OBJECTIVE: We aimed to clarify factors which affect the postoperative femoral anteroposterior position relative to the tibia under WB conditions (A-P position) in medial pivot (MP) TKA. METHODS: Among 126 knees (81 women) with primary TKA using MP prosthesis for varus osteoarthritic knees, 70 knees had cruciate-retaining inserts (CR) with the reduced conformity and 56 knees had cruciate-substituting inserts (CS) with the full conformity. Using the three-dimensional (3D) assessment system, the associations between the A-P position and the factors including pre- and postoperative lower extremity alignment, component positions, and posterior femoral condylar offset (PCO), were assessed regarding the type of inserts. RESULTS: Significant correlations were seen between the A-P position and posterior tibial slope (PTS), medial PCO, and lateral PCO. Regarding the difference between the two inserts, the PTS, medial PCO, and lateral PCO significantly correlated with the A-P position in the CR, but only the PTS correlated in the CS. CONCLUSIONS: The increased PTS and decreased PCO were the dominant factors for the A-P position in MP TKA.

Unidirectional output from a quantum-dot single-photon source hybrid integrated on silicon.

We report a quantum-dot single-photon source (QD SPS) hybrid integrated on a silicon waveguide embedding a photonic crystal mirror, which reflects photons and enables efficient unidirectional output from the waveguide. The silicon waveguide is constituted of a subwavelength grating so as to maintain the high efficiency even under the presence of stacking misalignment accompanied by hybrid integration processes. Experimentally, we assembled the hybrid photonic structure by transfer printing and demonstrated single-photon generation from a QD and its unidirectional output from the waveguide. These results point out a promising approach toward scalable integration of SPSs on silicon quantum photonics platforms.

Association between the toe angle and bony factors in the transverse plane for osteoarthritic knees compared with healthy knees.

BACKGROUND: The association between the toe angle and bony rotational factors is critical to explain issues related to the onset and progression of knee osteoarthritis (OA). OBJECTIVE: The study aimed to clarify the association between the toe angle and each of the femoral and tibial transvers direction relative to gait direction, rotational knee alignment, and bony torsional deformity for the subjects with knee OA. METHODS: This study evaluated 58 knees in 24 healthy elderly (72 ± 5 years) and 34 varus knee OA (72 ± 6 years). A three-dimensional (3D) assessment system was used on 3D models and biplanar long-leg radiographs with the toe angle reflecting gait direction, applying a 3D-to-2D image registration technique. The main parameters on the transverse plane were: (1) toe angle, (2) transverse direction of the femur and tibia relative to the gait direction, (3) femoral neck anteversion, (4) condylar twist angle, (5) tibial torsion, and (6) rotational knee alignment. RESULTS: The alignment parameters, except for the tibial transverse direction, were different between healthy and osteoarthritic knees. In knee OA, the femoral neck anteversion - femoral transverse direction (p = 0.001), femoral transverse direction - tibial transverse direction (p < 0.001), and tibial transverse direction - toe angle (p < 0.001) were associated. CONCLUSIONS: The osteoarthritic knees showed that the femoral neck anteversion was associated with the femoral transverse direction, which determined the tibial transverse direction by adjusting the rotational knee alignment, leading to the toe angle.

Experimental demonstration of topological slow light waveguides in valley photonic crystals.

We experimentally demonstrate topological slow light waveguides in valley photonic crystals (VPhCs). We employed a bearded interface formed between two topologically-distinct VPhCs patterned in an air-bridged silicon slab. The interface supports both topological and non-topological slow light modes below the light line. By means of optical microscopy, we observed light propagation in the topological mode in the slow light regime with a group index ng over 30. Furthermore, we confirmed light transmission via the slow light mode even under the presence of sharp waveguide bends. In comparison between the topological and non-topological modes, we found that the topological mode exhibits much more efficient waveguiding than the trivial one, demonstrating topological protection in the slow light regime. This work paves the way for exploring topological slow-light devices compatible with existing photonics technologies.

Influence of posterior tibial slope on three-dimensional femorotibial alignment under weight-bearing conditions in healthy Japanese elderly people.

BACKGROUND: Assessment of three-dimensional (3D) femorotibial alignment is essential for successful knee osteoarthritis treatment in the elderly. The complex morphology of the posterior tibial slope (PTS) might have an influence on sagittal and rotational alignment and the positional relationship between the femur and tibia in the anterior-posterior (AP) direction under weight-bearing conditions. OBJECTIVE: This study aimed to clarify the association between the PTS and 3D femorotibial alignment under weight-bearing conditions in healthy Japanese elderly individuals. METHODS: We investigated the 3D femorotibial alignment of 110 lower extremities of 55 healthy individuals (26 women, 29 men, mean age: 70 ± 6 years). Using our previously reported 3D-to-2D image registration technique, we evaluated the 3D hip-knee-ankle angle (3DHKA) in the sagittal plane, rotational alignment, and the distance between the femoral and tibial origins in the AP direction (tibial AP position) as femorotibial alignment parameters under weight-bearing conditions. We assessed the medial and lateral PTS and their angular difference (PTS difference) as PTS parameters. Stepwise multiple linear regression analysis was performed using PTS parameters and other possible confounders (age, sex, height, and weight) as the independent variables and femorotibial alignment parameters as the dependent variable. RESULTS: Weight (𝛽 = 0.393, p < 0.001) and lateral PTS (𝛽 = 0.298, p < 0.001) were the predictors associated with 3DHKA in the sagittal plane. Lateral PTS (𝛽 = 0.304, p = 0.001) was the only predictor associated with the tibial AP position. Sex (𝛽 = -0.282, p = 0.002) and PTS difference (𝛽 = -0.231, p = 0.012) were associated with rotational alignment. CONCLUSIONS: We found that a steeper lateral PTS resulted in a more flexed knee and anterior tibia. The PTS difference was positively correlated with tibial external rotation. Our data could be used as the standard reference for realignment surgery to ensure PTS is appropriately maintained.

New evaluation indices for rotational knee angles in standing anteroposterior knee radiographs.

BACKGROUND: Identifying the time course of rotational knee alignment is crucial for elucidating the etiology in knee osteoarthritis. OBJECTIVE: The aim of this study was to propose new rotational indices for calculating the change in relative rotational angles between the femur and tibia in standing anteroposterior (AP) radiographs. METHODS: Forty healthy elderly volunteers (20 women and 20 men; mean age, 70 ± 6 years) were assessed. The evaluation parameters were as follows: (1) femoral rotational index: the distance between the sphere center of the medial posterior femoral condyle and the lateral edge of the patella, and (2) tibial rotational index: the distance between the medial eminence of the tibia and the lateral edge of the fibula head. The indices were standardized by the diameter of the sphere of the medial posterior femoral condyle. This study (1) identified the relationship between changes in rotational indices and the simulated rotational knee angles in the standing position, (2) proposed a regression equation for the change in relative rotational angles between the femur and tibia in standing AP radiographs, and (3) verified the accuracy of the regression equation. RESULTS: The rotational indices increased in direct proportion to simulated rotational knee angles (femoral index: r > 0.9,p < 0.0001; tibial index: r > 0.9, p < 0.0001). Based on the results, the regression equation with the accuracy of 0.45 ± 0.26° was determined. CONCLUSIONS: The proposed regression equations can potentially predict the change in relative rotational angles between the femur and tibia in a pair of standing AP radiographs taken at different dates in longitudinal studies.

Cortical thickness of the tibial diaphysis reveals age- and sex-related characteristics between non-obese healthy young and elderly subjects depending on the tibial regions.

PURPOSE: This study aimed to evaluate the age- and sex-related characteristics in cortical thickness of the tibial diaphysis between non-obese healthy young and elderly subjects as reference data. METHODS: The study investigated 31 young subjects (12 men and 19 women; mean age, 25 ± 8 years) and 54 elderly subjects (29 men and 25 women; mean age, 70 ± 6 years). Three-dimensional estimated cortical thickness of the tibial diaphysis was automatically calculated for 5000-9000 measurement points using the high-resolution cortical thickness measurement from clinical computed tomography data. In 12 assessment regions created by combining three heights (proximal, central, and distal diaphysis) and four areas of the axial plane at 90° (medial, anterior, lateral, and posterior areas) in the tibial coordinate system, the standardized thickness was assessed using the tibial length. RESULTS: As structural characteristics, there were no differences in the medial and lateral thicknesses, while the anterior thickness was greater than the posterior thickness in all groups. The sex-related difference was not shown. As an age-related difference, elderly subjects showed greater or lesser cortical thickness than the young subjects, depending on the regions of the tibia. CONCLUSIONS: Cortical thickness was different depending on sex, age, and regions in the tibia. The results of this study are of clinical relevance as reference points to clarify the causes of various pathological conditions for diseases. LEVEL OF EVIDENCE: Level 3.

The majority of patellar avulsion fractures in first-time acute patellar dislocations included the inferomedial patellar border that was different from the medial patellofemoral ligament attachment.

PURPOSE: Accurate assessment of the locations of patellar avulsion fractures in acute patellar dislocations is clinically relevant for decision making for treatment. The study aim was to classify the locations of patellar avulsion fractures with a focus on the ligament attachments of medial stabilizing structures. METHODS: Out of 131 first-time acute traumatic patellar dislocations, 61 patients had patellar fractures. Subsequently, 10 patients with isolated osteochondral fractures of the articular surface in the patella were excluded. Finally, 51 patients (34 females and 17 males, average age: 18.5 years, 95% CI 16.1-20.9) were included in the study cohort. Based on the locations of the patellar attachment, the patients were divided into three groups: the superior group [medial patellofemoral ligament (MPFL) attachment], inferior group [medial patellotibial ligament (MPTL)/medial patellomeniscal ligament (MPML) attachment], and mixed group. RESULTS: In the patellar avulsion group (51 patients), the superior group, mixed group, and inferior group contained 8/51 (16%), 12/51 (24%), and 31/51 (61%) patients, respectively. CONCLUSIONS: This study showed that 84% of the patellar avulsion fractures were located in the inferomedial patellar border, which consisted of MPTL/MPML attachments that were clearly different from the true "MPFL" attachment at the superomedial patellar border. In terms of the clinical relevance, the acute surgical repair of MPTL/MPML attachments in the inferomedial patellar border may not sufficiently control the patella if optimal management of the MPFL is not performed. LEVEL OF EVIDENCE: IV.

Affinity shift of ATP upon glycerol binding to a glycerol kinase from the hyperthermophilic archaeon Thermococcus kodakarensis KOD1.

Glycerol kinase (GK) is a key enzyme of glycerol metabolism. It participates in glycolysis and lipid membrane biosynthesis. A hexamer of GK from the hyperthermophilic archaeon Thermococcus kodakarensis KOD1(Tk-GK) was identified as a substrate-binding form of the enzyme. Here, the X-ray crystal structure analysis and the biochemical analysis was done and the relationships between its unique oligomer structure and substrate binding affinity were investigated. Wild type GK and mutant K271E GK, which disrupts the hexamer formation interface, were crystallized with and without their substrates and analyzed at 2.19-3.05 Å resolution. In the absence of glycerol, Tk-GK was a dimer in solution. In the presence of its glycerol substrate, however, it became a hexamer consisting of three symmetrical dimers about the threefold axis. Through glycerol binding, all Tk-GK molecules in the hexamer were in closed form as a result of domain-motion. The closed form of Tk-GK had tenfold higher ATP affinity than the open form of Tk-GK. The hexamer structure stabilized the closed conformation and enhanced ATP binding affinity when the GK was bound to glycerol. This molecular mechanism is quite simple activity regulation mechanism among known GKs.

The inclination of the femoral medial posterior condyle was almost vertical and that of the lateral was tilted medially.

PURPOSE: The purpose of this study was to three-dimensionally analyse the size and shape of the femoral posterior condyles of the normal knee. METHODS: A total of 62 healthy Japanese volunteers (37 males and 25 females) providing a sample of 124 normal knee joints, who had no knee-related symptoms and no history of major trauma, underwent computed tomography scans of the bilateral femur and tibia. Three-dimensional digital models of the femur were constructed from computed tomography data using visualisation and modelling software. The following parameters were evaluated: (1) the radii of the posterior condyles approximated to spheres and (2) the inclination angle of the posterior condyles in the coronal plane of the femoral coordinate system. RESULTS: The radii of the medial and lateral condyles approximated to spheres were 17.0 ± 1.6 and 17.1 ± 1.8 mm, respectively and were not different. The inclination angles of the medial and lateral condyles in the coronal plane were - 0.6° ± 4.6° and 9.7° ± 5.7°, respectively. The medial condyle was almost vertical, whereas the lateral one was medially tilted. CONCLUSIONS: This study found an asymmetrical inclination between medial and lateral condyles. This may be related to the asymmetrical motion of the knee, which is known as medial pivot motion. This finding provides valuable morphological information and may be useful for implant designs for total knee arthroplasty. LEVEL OF EVIDENCE: IV.

Age- and sex-related characteristics in cortical thickness of femoral diaphysis for young and elderly subjects.

INTRODUCTION: Cortical thickness of the femoral diaphysis is assumed to be a preferred parameter in the assessment of the structural adaptation by mechanical use and biological factors. This study aimed to investigate the age- and sex-specific characteristics in cortical thickness of the femoral diaphysis between young and elderly non-obese people. MATERIALS AND METHODS: This study investigated 34 young subjects (21 men and 13 women; mean age: 27 ± 8 years) and 52 elderly subjects (29 men and 23 women; mean age: 70 ± 6 years). Three-dimensional (3D) cortical thickness of the femoral diaphysis was automatically calculated for 5000-8000 measurement points using the high-resolution cortical thickness measurement from clinical CT data. In 12 assessment regions created by combining three heights (proximal, central, and distal diaphysis) and four areas of the axial plane at 90° (medial, anterior, lateral, and posterior areas) in the femoral coordinate system, the standardized thickness was assessed using the femoral length. RESULTS: As per the trends, (1) there were no differences in medial and lateral thicknesses, while the posterior thickness was greater than the anterior thickness, (2) the thickness in men was higher than that in women, and (3) the thickness in young subjects was higher than that in elderly subjects. CONCLUSIONS: The results of this study are of clinical relevance as reference points to clarify the causes of various pathological conditions for diseases of the lower extremities.

Articular surface of the medial proximal tibia is aligned parallel to the ground in three-dimensional space under weight-bearing conditions in healthy and varus osteoarthritic knees.

PURPOSE: To test the hypothesis that an inclined articular surface on the medial proximal tibia is aligned more parallel to the ground in three-dimensional (3D) space under weight-bearing (WB) conditions (parallel phenomenon) than under non-WB (NWB) conditions in healthy and varus osteoarthritic knees. METHODS: We examined 55 healthy knees (26 women, 29 men; mean age, 70 ± 6 years) and 108 varus osteoarthritic knees (66 women, 16 men; mean age, 74 ± 7 years). For the evaluation under WB conditions, a 3D assessment system was used on biplanar long-leg radiographs and 3D bone models using a 3D-to-2D image registration technique. In addition, the least square method was used to determine the approximation plane. The angles between the normal vector for the approximation plane of an articular surface on the medial proximal tibia and each axis of the tibial or world coordinate system were calculated. RESULTS: Morphologically, the inclination of the approximation plane was steeper in osteoarthritic knees than in healthy knees (p < 0.0001). The approximation plane was aligned more parallel to the ground under WB conditions than under NWB conditions in healthy (p < 0.0001) and osteoarthritic knees (p < 0.0001). CONCLUSIONS: The parallel phenomenon in the medial proximal tibia was confirmed for healthy and varus osteoarthritic knees. The medial proximal tibia plays an important role in the parallel phenomenon, assumingly associated with varus alignment and varus thrust. The inclination of the medial proximal tibia may become a new parameter for imaging investigations. LEVEL OF EVIDENCE: III.

Correlation between posterior tibial slope and sagittal alignment under weight-bearing conditions in osteoarthritic knees.

INTRODUCTION: Posterior tibial slope (PTS) and sagittal alignment are important factors in the etiology of knee osteoarthritis and knee surgery. Clinically, sagittal alignment, which indicates flexion contracture of the knee, contributes to knee function in weight-bearing (WB) conditions. PTS and sagittal alignment under WB conditions in varus osteoarthritic knees are presumed to affect each other, but their association remains unclear. In this study, we aimed to clarify the association. MATERIAL AND METHODS: In total, 140 osteoarthritic varus knees were investigated. Under WB conditions, a three-dimensional (3D) alignment assessment system was applied via biplanar long-leg X-rays, using 3D-to-2D image registration technique. The evaluation parameters were as follows: 1) 3D mechanical flexion angle (3DMFA) in regards to sagittal alignment, 2) passing point in the WB line (PP), and 3) medial and lateral PTS. RESULTS: The medial and lateral PTS showed a positive correlation with 3DMFA and PP, respectively (medial PTS-3DMFA, p = 0.001; medial PTS-PP, p < 0.0001; lateral PTS-3DMFA, p < 0.0001; lateral PTS-PP, p = 0.002). The flexion contracture group with 3DMFA >5° demonstrated greater PTS than non-flexion contracture group (medial PTS, p = 0.006; lateral PTS, p = 0.006). CONCLUSIONS: Both medial and lateral PTS were correlated with sagittal alignment under WB conditions and were larger in the flexion contracture group. This finding can explain the function to take the load articular surface parallel to the ground for holding the balance in WB conditions in the sagittal plane for osteoarthritic knees. Moreover, surgeons may be required to decrease the PTS during knee arthroplasty to restore full extension in knees of patients with fixed flexion contracture.

Contribution of sex and body constitution to three-dimensional lower extremity alignment for healthy, elderly, non-obese humans in a Japanese population.

BACKGROUND: Humans support their bodies exclusively by vertical balance in bipedal locomotion, and the body, especially the lower extremity, generally changes with age. Sex and body constitution are assumed to be associated with lower extremity alignment, but this association remains to be elucidated. This study sought to clarify this association in healthy, elderly, non-obese humans in a Japanese population. METHODS: The present study investigated 55 healthy volunteers (mean age: 70 ± 6 years). A 3D extremity alignment system was applied under weight-bearing conditions on biplane long lower extremities X-rays using a 3D-to-2D image registration technique. The evaluation parameters included 3D hip-knee-ankle angle (3DHKA) alignment in the coronal (coronal alignment) and sagittal planes (sagittal alignment) and rotational alignment between the femur and tibia. The influences of sex and body constitution on all the alignment were analyzed. RESULTS: Multiple linear regression analysis with the dependent variable of each alignment showed that sex was the dominant factor for coronal and rotational alignment (coronal: p <  0.01; rotational: p <  0.01), and body weight was the dominant factor for sagittal alignment (p <  0.01). CONCLUSIONS: The association of sex with coronal and rotational alignment and of body constitution with sagittal alignment were proved in healthy, elderly, non-obese humans in a Japanese population. This finding can lead to further understanding of the etiology of many diseases and age-related changes.

Crystal structure of highly thermostable glycerol kinase from a hyperthermophilic archaeon in a dimeric form.

The crystal structure of glycerol kinase from the hyperthermophilic archaeon Thermococcus kodakaraensis (Tk-GK) in a dimeric form was determined at a resolution of 2.4 A. This is the first crystal structure of a hyperthermophilic glycerol kinase. The overall structure of the Tk-GK dimer is very similar to that of the Escherichia coli glycerol kinase (Ec-GK) dimer. However, two dimers of Ec-GK can associate into a tetramer with a twofold axis, whereas those of Tk-GK cannot. This may be the reason why Tk-GK is not inhibited by fructose 1,6-bisphosphate, because the fructose 1,6-bisphosphate binding site is produced only when a tetrameric structure is formed. Differential scanning calorimetry analyses indicate that Tk-GK is a highly thermostable protein with a melting temperature (T(m)) of 105.4 degrees C for the major transition. This value is higher than that of Ec-GK by 34.1 degrees C. Comparison of the crystal structures of Tk-GK and Ec-GK indicate that there is a marked difference in the number of ion pairs in the alpha16 helix. Four ion pairs, termed IP1-IP4, are formed in this helix in the Tk-GK structure. To examine whether these ion pairs contribute to the stabilization of Tk-GK, four Tk-GK and four Ec-GK derivatives with reciprocal mutations at the IP1-IP4 sites were constructed. The determination of their stabilities indicates that the removal of each ion pair does not affect the stability of Tk-GK significantly, whereas the mutations designed to introduce one of these ion pairs stabilize or destabilize Ec-GK considerably. These results suggest that the ion pairs in the alpha16 helix contribute to the stabilization of Tk-GK in a cooperative manner.

Crystallization and preliminary X-ray diffraction study of glycerol kinase from the hyperthermophilic archaeon Thermococcus kodakaraensis.

Glycerol kinase from the hyperthermophilic archaeon Thermococcus kodakaraensis was crystallized and preliminary crystallographic studies of the crystals were performed. Crystals were grown at 293 K by the sitting-drop vapour-diffusion method. Native X-ray diffraction data were collected to 2.4 A resolution using synchrotron radiation at station BL44XU of SPring-8. The crystal belongs to the rhombohedral space group R3, with unit-cell parameters a = b = 217.48, c = 66.48 A. Assuming the presence of two molecules in the asymmetric unit, the V(M) value was 2.7 A(3) Da(-1) and the solvent content was 54.1%. The protein was also cocrystallized with substrates and diffraction data were collected to 2.7 A resolution.