Biomedical subjects

S Ando

Publications and source records attributed to S Ando.

At least 145 records · Page 8Linked to original sources

PubMed1996

Enhancement of smooth muscle contraction with protein phosphatase inhibitor 1: activation of inhibitor 1 by cGMP-dependent protein kinase.

New method for purification of phosphatase inhibitor 1 (PPI-1) was developed which avoid the phosphorylation of PPI-1 during the purification and provides a high yield of highly pure preparation. Using this preparation, it was shown that PPI-1 was stoichiometrically phosphorylated by cGMP-dependent protein kinase at Thr-35 and the phosphorylated PPI-1 potently inhibited protein phosphatase 1. Addition of the phosphorylated PPI-1 to beta-escin-skinned single smooth muscle cells resulted in force development of the cells at the submaximal pCa2+. The results suggest that the phosphorylation of PPI-1 can be the mechanism for modifying the Ca2+ sensitivity of smooth muscle contractile response.

Amino Acid Sequence↗

PubMed1996

Impaired synaptic functions with aging as characterized by decreased calcium influx and acetylcholine release.

Age-related alterations of presynaptic functions were studied in terms of acetylcholine (ACh) synthesis and release using synaptosomes isolated from mouse brain cortices. The following three findings were obtained: 1) Choline acetyltransferase activity and ACh production rate remained constant throughout all ages tested. This observation, obtained with synaptosomes, was not consistent with data reported for brain slices (Gibson GE, Peterson C: J Neurochem 37:978-984, 1981). Various conditions, such as low glucose or membrane depolarization, modulated ACh synthesis to similar extents in young and aged synaptosomes. 2) Depolarization-induced release of ACh from synaptosomes significantly decreased in the senescent stage. The fraction of ACh released from aged synaptosomes was less than that released from young synaptosomes, although the ACh contents in the synaptosomes did not change with age. 3) Calcium influx induced by depolarization was lower in the synaptosomal preparations from aged mice than in those from young mice. A strong positive correlation was observed between the amounts of ACh released and the increased calcium levels when the data for all preparations, both from young and aged mice, were plotted. This indicates that diminished calcium influx may cause the reduced ACh release by aged synapses. The present study provides evidence for an age-related decrease in presynaptic functions, that is, a reduction in calcium influx via voltage-dependent calcium channels followed by a decreased ACh release from synapses despite an abundance of ACh within the synapses.

Acetylcholine↗

PubMed1996

An epitope on Ki antigen recognized by autoantibodies from lupus patients shows homology with the SV40 large T antigen nuclear localization signal.

OBJECTIVE: Epitopes on Ki antigen were analyzed using synthetic peptides, including KILT, a 16-mer peptide with an amino acid sequence homologous to the SV40 large T antigen nuclear localization signal (SV40 T NLS). METHODS: In addition to KILT, 4 synthetic peptides, all potential epitopes on Ki antigen according to computer analysis, were prepared and tested for reactivity with 49 anti-Ki-positive lupus sera by enzyme-linked immunosorbent assay. RESULTS: Eighteen sera reacted with KILT, but not with other peptides. The reaction of anit-Ki sera with KILT was specifically inhibited by recombinant Ki antigen. Eight of 49 anti-Ki sera reacted with a 7-mer synthetic peptide of SV40 T NLS, and the reaction was specifically inhibited by KILT. CONCLUSION: The 16-mer Ki peptide containing the sequence homologous to the SV40 T NLS is one of the antigenic epitopes recognized by anti-Ki antibodies in lupus sera.

Amino Acid Sequence↗

PubMed1996

Modulation of cholinergic synaptic functions by sialylcholesterol.

The effects of sialylcholesterol, a synthetic ganglioside analogue, on cholinergic synaptic functions were investigated using synaptosomes prepared from C57BL/6 mouse brain cortices. Addition of alpha-sialylcholesterol stimulated high K (50 mM)-evoked acetylcholine (ACh) release from synaptosomes at concentrations ranging from 1 to 5 microM. The beta-anomer of the sialyl compound also increased the neurotransmitter release at 5 microM, but the effect was much smaller than that of the alpha-anomer. Beta-sialylcholesterol appeared to increase high-affinity choline uptake and Ach synthesis, resulting in an increment in the release of ACh. On the other hand, alpha-sialylcholesterol did not change the synthetic rate of ACh, and instead it increased the depolarization=induced influx of calcium ions into synaptosomes, while the beta-anomer did not affect the divalent cation influx. The enhanced calcium influx is thought to increase ACh release from synaptosomes treated with alpha-sialylcholesterol. These results imply that the two anomers of sialylcholesterol may modulate the synaptic membrane machinery differently, that is, the alpha-anomer may activate voltage-dependent calcium channels and the beta-anomer may facilitate high-affinity choline uptake. In order to evaluate the ameliorating effect of sialylcholesterol, alpha-sialylcholesterol was applied to the synaptosomes from aged mice (34 months old), which have been shown to have a decreased ACh release (Tanaka et al., 1995, J Neurosci Res, in press [1]). The reduced neurotransmitter release recovered to the levels of younger animals, suggesting that sialylcholesterol might have a potential therapeutic use for restoring synaptic function that occurs in aged brains.

Acetylcholine↗

PubMed1996

The direct effect of interferon-gamma on human eosinophilic leukemia cell lines: the induction of interleukin-5 mRNA and the presence of an interferon-gamma receptor.

The EoL-1 and EoL-3, human eosinophilic leukemia cell lines, have been used as models for studying the maturation and the function of human eosinophils. We investigated the effects of interferon-gamma (IFN-gamma) on superoxide anion (O2-) production of these cell lines and interleukin-5 (IL-5) mRNA expression in the EoL-1. O2- was measured by chemiluminescence of MCLA, one of cypridina luciferin analogs. The O2- production of fMLP-stimulated EoL-1 and EoL-3 was increased by the IFN-gamma treatment. IL-5 mRNA expression was detected in the IFN-gamma-treated EoL-1 by reverse transcription-polymerase chain reaction (RT-PCR). Further, we examined IFN-gamma receptor 1 mRNA expression in these cell lines and peripheral blood eosinophils by means of northern blot hybridization. IFN-gamma receptor 1 mRNA was detected in the EoL-3 and the IFN-gamma-treated EoL-1. A weak expression of IFN-gamma receptor 1 mRNA was detected in peripheral blood eosinophils isolated from a patient with eosinophilia. These results suggest that IFN-gamma may act on eosinophils directly through its receptor.

Antigens, CD↗

PubMed1996

A novel ganglioside, 9-O-acetyl GD1b, is recognized by serum antibodies in Guillain-Barré syndrome.

A hitherto undescribed ganglioside was detected in a crude ganglioside fraction of bovine brain using an IgM M-protein binding to Gal beta 1, 3GalNAc residue. We purified and identified it as 9-O-acetyl GD1b based on results of alkali treatment that yielded GD1b and results of fast atom bombardment-mass and gas chromatography-mass spectrometries. 9-O-acetyl GD1b was also found to be present in human peripheral nerve tissue. The reactivities of the serum antibodies from patients with Guillain-Barré syndrome to 9-O-acetyl GD1b, GD1b, and GM1 were determined by ELISA and TLC immunostaining. Nineteen of 85 serum samples from Guillain-Barré syndrome patients had antibodies that bound to 9-O-acetyl GD1b: 14 of the positive samples also reacted with GM1 and GD1b, three reacted with GM1 but not with GD1b, one with GD1b but not with GM1, and one with neither GM1 nor GD1b. These results show that a subset of patients with Guillain-Barré syndrome had antibodies that react with 9-O-acetyl GD1b; therefore, this ganglioside can serve as a target antigen against the antibodies present in Guillain-Barré syndrome.

Animals↗

PubMed1996

Role of calmodulin-dependent protein kinase II in the acute stimulation of aldosterone production.

Acute aldosterone production in adrenocortical cells is highly dependent on calcium (Ca2+) and calmodulin (CaM) activation. To determine the role of calmodulin-dependent protein kinase II (CaM kinase II) in human adrenal aldosterone production, the action of KN93 (a specific CaM kinase II inhibitor) on human adrenocortical H295R cells was examined. The stimulation of aldosterone, production by angiotensin II (Ang II) and potassium (K+) were inhibited by KN93 in a concentration-dependent manner with an IC50 of approximately 0.9 and approximately 0.5 microM, respectively. Aldosterone production was also stimulated by treatment with the calcium channel activator Bay K 8644 (Bay K) (1 microM). This production was inhibited in a concentration-dependent manner by KN93 with an IC50 of between 1 and 3 microM. No inhibition by KN93 (0.3-3 microM) or by the calmodulin inhibitor calmidazolium (0.03-0.3 microM) was observed for 22R-hydroxycholesterol (22R-OHChol) stimulation of aldosterone production. Because 22R-OHChol is a substrate for the cytochrome P450 cholesterol side-chain cleavage enzyme (P450scc) and does not require active transport to the mitochondria, these results indicate that KN93 does not directly inhibit P450scc or later steps leading to aldosterone synthesis. To investigate the site of KN93 action further we examined its effect on agonists induction of steroidogenic acute regulatory (StAR) protein, which was recently shown to regulate the movement of cholesterol from the outer to the inner mitochondrial membranes. Induction of StAR protein in H295R cells by Ang II, or Bay K was not affected by co-treatment with KN93 at concentration which blocked steroidogenesis by 60-80%. These results indicate a direct role of CaM kinase II in Ang II and K+ simulation of aldosterone production and support the hypothesis that CaM kinase II may be involved in the process of cholesterol mobilization to the mitochondria.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗

PubMed1996

Application of a newly developed 3-D deformation measurement system to prosthetic dentistry.

A new system for measuring three-dimensional deformation has been developed. The main components of the system are a fixed TV camera and vertically scanning projectors. The advantages of the system are: (1) since there is no mechanical contact between the object and the sensing unit, hardness and form of the object do not have to be considered; (2) the system does not need any precise geometry except for the reproducibility of the vertical scanning stage; (3) the time required for one measurements is extremely short; and (4) the system is not expensive. From the basic experiments for testing accuracy, this system has about +/- 30 microns range deviation against 1000 microns deformation of the object. The system has been applied to investigate the influence of various impression procedures for a removable partial denture on the displacement of the simulated soft mucosal tissue. It is revealed that deformation forms are expressed precisely. Our results suggest that this measurement system is quite useful for research in dentistry and that it could be widely applied.

Biomechanical Phenomena↗

PubMed1996

Effects of asymmetric dynamic and isometric liftings on strength/force and rating of perceived exertion.

A laboratory study was undertaken to determine the postural and physical characteristics and subjective stress during dynamic lifting of a usual load (10 kg) compared with during isometric lifting. The authors also aimed to clarify the effects of asymmetric lifting on these parameters. The subjects were thirteen male college students. They were asked to lift a box weighing 10 kg. They performed sixteen different lifting tasks from the floor to a height of 71 cm, involving a combination of three independent factors: two lifting modes (isometric lifting and dynamic lifting), four lifting angles in relation to the sagittal plane (sagittal plane, right 45 degree, right 90 degree and left 90 degree planes) and two lifting postures (squat and stoop). For each lifting task, strengths or forces and ground reaction forces were measured. At the end of each task, the authors asked the subjects to rate their perceived exertion (RPE) during lifting at ten sites of the body. Angle factor had a significant effect on isometric strengths and dynamic peak forces. Isometric strengths during the maximum 3 s were highest in lifting in the right 45 degree plane, followed by that in the sagittal plane, while those in the right 90 degree and left 90 degree planes were the lowest. However, peak forces in dynamic lifting were the highest in the lifting in the sagittal plane, followed by that in the right 45 degree plane, while those in the right 90 degree and left 90 degree planes were the lowest. Postural factor had a significant effect on height at peak force, which is higher in squat lifting than in stoop lifting. RPEs for the left arm, the backs and the right whole body in isometric lifting were significantly higher than in dynamic lifting of 10 kg. There were remarkably high RPEs for the ipsilateral thigh to the box in right 90 degree and left 90 degree planes during both isometric and dynamic liftings. Locations of the resultant force consisting of three component forces on the force plate were closer to the foot on the same side as the box in asymmetric lifting. Thus, some similarities and differences were found between isometric lifting and dynamic liftings regarding the indexes of strength used in this experiment. The authors consider that the subjects used the foot nearer to the box as a fulcrum during asymmetric lifting. Dynamic measurement using the 10 kg weight is less stressful than the conventional isometric measurement. It was possible to obtain the height data at peak force and time-based changes in the force and the box location during lifting only through dynamic lifting measurement. The results provide new knowledge about the biomechanical features of dynamic lifting tasks.

Adolescent↗

PubMed1996

[Comparative susceptibility of 10 antimicrobials against Pseudomonas aeruginosa isolated from complicated urinary tract infection on 1992 & 1993 vs. 1994 & 1995].

The susceptibility of 10 antimicrobials against P. aeruginosa isolated from complicated UTI during 1992-1995 was determined, and the yearly trend was examined. The drug tested included 3 cephems (ceftazidime [CAZ], cefpirome [CPR], cefclidin [CFCL]), 1 monobactam (aztreonam [AZT]), 2 carbapenem (imipenem/cilastatin [IPM/CS], biapenem [BIPM]), 2 aminoglycosides (netilmicin [NTL], gentamicin [GM]) and 2 new quinolones (ofloxacin [OFLX], ciprofloxacin [CPFX]). A total number of isolates of which MIC were determined was 77 in 1992/1993 and 70 in 1994/1995. MIC50/MIC90(micrograms/ml) on the isolates were as follows (1992/1993 1994/1995); 3.13/ 100 12.5/50 in CAZ, 12.5/100 12.5/100 in CPR, 3.13/25 1.56/25 in CFCL, 6.25/50 12.5/100 in AZT, 6.25/25 3.13/25 in IPM/CS, 1.56/6.25 0.78/50 in BIPM, 12.5/100 6.25/100 in NTL, 6.25/ 50 6.25/100 < in GM, 25.100 < 25/100 < in OFLX, 6.25/100 6.25/100 < in CPFX. When the susceptibility of the yearly trend is compared, no significant changes were detected among the drugs tested expect the decrease of susceptibility on CAZ, AZT and increase of it on IPM/CS. From the data obtained, against complicated UTI infected by P. aerugunosa, CFCL, IPM/CS and BIPM are considered to drugs of first choice in the treatment.

Anti-Bacterial Agents↗

PubMed1996

Genetic variability of the common nod gene in soybean bradyrhizobia isolated in Thailand and Japan.

To determine the taxonomic relationship between Thai soybean bradyrhizobia and soybean bradyrhizobia from other regions, a total of 62 Bradyrhizobium strains were isolated in Thailand. The genetic diversity of the strains was examined with reference to 46 Japanese and 15 USDA strains. The degree of sequence divergence in and around common nod gene regions of the 123 strains was estimated by restriction fragment length polymorphism analysis using the Bradyrhizobium japonicum USDA 110 common nodDYABC gene probe. The phylogenetic grouping of the strains resulted in four major clusters. Cluster 1 comprised the Japanese and USDA strains, which originated in temperate regions, whereas clusters 3 and 4 comprised the tropical Thai strains. Cluster 1 strains comprised the DNA homology groups I and Ia, and hence, were classified as B. japonicum. Cluster 2 strains were in the DNA homology group II, and hence, were classified as Bradyrhizobium elkanii. Clusters 3 and 4 strains, however, did not correspond to any known DNA homology groups. These results indicate that Thai soybean bradyrhizobia are distantly related to B. japonicum and B. elkanii.

Bacterial Proteins↗

PubMed1996

Transfer function analysis of central arc of aortic baroreceptor reflex in rabbits.

While electrically stimulating the aortic depressor nerve (ADN) pseudorandomly, we recorded renal sympathetic nerve activity (RSNA) and systemic arterial pressure (SAP) in 19 alpha-chloralose-anesthetized rabbits with sinoaortic denervation. From the recorded signals, we determined the transfer functions from ADN stimulation by a pseudorandom binary sequence to RSNA [HCMD.RSNA(f)] and to SAP [HCMD.SAP(f)]. The modulus of HCMD.RSNA(f) was flat over 0.0122-0.8 Hz, whereas the phase lag increased linearly with frequency. Thus the central transduction appeared not to modify the relative amplitude of the signals from the baroreflex afferents but to provide a fixed time delay (approximately 400 ms). In contrast, the modulus of HCMD.SAP(f) decreased precipitously toward high frequencies, and the degree of the phase lag was larger than that of HCMD.RSNA(f). We conclude that 1) the transfer property of the central are does not significantly modify the relative amplitude of the frequency components of the baroreflex afferents but provides a fixed time delay and 2) the frequency independence of the modulus of the transfer property is not preserved when the analysis is extended to SAP.

Animals↗

PubMed1996

Cycle length of periodic breathing in patients with and without heart failure.

Because apnea length during periodic breathing varies according to the preceding increase in ventilation and reduction in PaCO 2, differences in the cycle length of periodic breathing among patients with normal and impaired cardiac function might be explained by the influence of lung-to-carotid body circulatory delay, as reflected by lung-to-ear circulation time (LECT), on hyperpnea length rather than on apnea length. It was therefore hypothesized that circulatory delay is an important determinant of periodic-breathing hyperpnea length but not apnea length. To test this hypothesis, LECT, periodic-breathing cycle length, apnea length, and hyperpnea length were compared in 10 patients with idiopathic central sleep apnea (ICSA), whose cardiac function was normal, as opposed to 10 with Cheyne-Stokes respiration and central sleep apnea (CSR-CSA) in association with congestive heart failure (CHF). As compared with ICSA patients, cycle length was significantly longer in patients with CSR-CSA (37.3 +/- 3.0 s versus 59.0 +/- 4.9 s, p < 0.005). This difference was due to significantly longer hyperpnea length in the CSR-CSA patients (16.7 +/- 2.8 s versus 36.7 +/- 3.4 s, p < 0.001), since apnea length was similar in the two groups. In addition, LECT was longer in the CSR-CSA patients (24.3 +/- 2.0 s versus 10.3 +/- 1.0 s, p < 0.001), and correlated strongly with cycle length (r = 0.88, p < 0.001) and hyperpnea length (r = 0.90, p < 0.001) but not with apnea length. LECT correlated inversely with cardiac output (r = -0.72, p < 0.006), indicating that LECT is a valid measure of circulatory delay. Thus, circulatory delay is an important determinant of hyperpnea length but not of apnea length in patients with ICSA and CSR-CSA.

Cardiac Output↗

PubMed1996

Vasospastic angina in patients with systemic triglyceride storage disease with Jordans' anomaly and cardiomyopathy.

Vasospastic angina was demonstrated clinically and angiographically in a 54-year-old patient with systemic triglyceride storage disease and cardiomyopathy. He and his younger sister had been diagnosed in 1985 as having systemic triglyceride storage with Jordans' anomaly. In 1993, he began complaining of rest and effort chest pain in the morning, which was accompanied by ST depression by ECG. Sublingual nitroglycerine was effective for treating this pain. Intracoronary injection of acetylcholine induced severe coronary vasoconstriction in the left anterior descending artery. Left ventricular contraction was diffusely impaired. Deposits of numerous triglyceride droplets and a decrease in the density of myofibrils in cardiocytes were found in the specimens obtained by endomyocardial biopsy. The impaired left ventricular contraction may have been due to the changes in myocardial cells. His sister complained of a similar chest pain that was completely controlled by calcium channel antagonis. Thus, our cases demonstrated vasospastic angina associated with cardiomyopathy secondary to systemic triglyceride storage disease with Jordans' anomaly, though the causal relationship between these conditions remains unclear.

Adult↗

PubMed1996

Phylogenetic relationships of species of the genus Kluyveromyces van der Walt (saccharomycetaceae) deduced from the partial sequences of 18S and 26S ribosomal RNAs.

In order to clarify the phylogenetic relationships of the species classified in the genus Kluyveromyces (Saccharomycetaceae), three partial base sequences of 18S and 26S rRNAs of eighteen strains were determined. The regions determined of the strains corresponded to positions 1451 through 1618 (168 bases) of 18S rRNA and to positions 1611 through 1835 (225 bases) and 493 through 622 (130 bases) of a strain (IFO 2376) of Saccharomyces cerevisiae. The analyses of the partial base sequences suggested that the genus Kluyveromyces is phylogenetically heterogeneous, ranging from the strains that are quite close to the strain of S. cerevisiae to the strains that are distinct enough to be classified in genera separate from the genus Saccharomyces. From our sequence data, we concluded that the extent of the genus Kluyveromyces should be restricted to only one species, K. polysporus, the type species of the genus. Kluyveromyces phaffii was also distinct enough to deserve another genus. Kluyveromyces cellobiovorus was not close to any of the strains of Kluyveromyces species examined, and should be excluded from the genus. Most of the strains of the species examined were fairly close to the strain of S. cerevisiae.

Base Sequence↗

PubMed1996

Phylogenetic relationships of species of the genus Saccharomyces Meyen ex Reess deduced from partial base sequences of 18S and 26S ribosomal RNAs (saccharomycetaceae).

In order to clarify the phylogenetic relationships among the yeast species classified in the genus Saccharomyces, partial base sequences of 18S and 26S ribosomal RNAs were determined for ten selected strains. The regions determined correspond to positions 1451 through 1618 of the 18S rRNA and positions 493 through 622 and 1611 through 1835 of the 26S rRNA in S. cerevisiae. Analyses of these partial base sequences suggested that the genus Saccharomyces is phylogenetically heterogeneous. Saccharomyces servazzii and S. unisporus showed identical or very similar sequences in all the three regions, and their phylogenetic distance from S. cerevisiae was large enough to introduce a genus independent of Saccharomyces. Saccharomyces kluyveri is also distant from all the other Saccharomyces species examined, and is likely to deserve a new genus. Estimated phylogenetic relationships between Saccharomyces and other genera characterized by the Q-6 system, such as species of Zygosaccharomyces. Torulaspora, Kluyveromyces, Arxiozyma, Pachytichospora, Nadsonia. Hanseniaspora, Kloeckeraspora, and Saccharomycodes, are also discussed.

Base Sequence↗

PubMed1996

Factors affecting induction of neurological disorders in mice by paralysis-inducing Friend-related PVC viruses.

Our previous studies showed that the passage of the Friend virus complex through rats generated variant MuLVs, designated PVC111, PVC211, PVC321 and PVC441, that induced neurological disorders associated with tremor and paralysis. In this study, we tested the pathogenicity of four different PVC viruses in mice. Although histopathological studies revealed spongiform degeneration in the spinal cords of NFS mice infected with each PVC virus, only PVC441 frequently induced tremor and paralysis. After a long latency, all of these viruses induced leukemia associated with severe anemia. Further studies with PVC441 revealed dose- and age-dependence for tremor induction. In contrast to NFS mice, BALB/c, DBA/2 and C57BL/6 mice infected with PVC441 virus showed no neurological symptoms, although the virus could be isolated from the tissues of central nervous system. Despite the absence of neurological symptoms, a high degree of neuronal degeneration in the lumbar spinal cord was found in PVC441-infected BALB/c mice. A low degree of neuronal degeneration was found in PVC441-infected DBA/2 or C57BL/6 mice. Genetic crosses of these resistant mice with susceptible NFS mice indicated that resistance to tremor induction by PVC441 was dominant in all mouse strains and suggested that various host genes may control the susceptibility of mice to tremor induction by PVC441 virus.

Animals↗

PubMed1996

Introducing a point mutation identified in a patient with pituitary resistance to thyroid hormone (Arg 338 to Trp) into other mutant thyroid hormone receptors weakens their dominant negative activities.

Clinical resistance to thyroid hormone (RTH) has been classified into generalized resistance to thyroid hormone (GRTH) and pituitary resistance to thyroid hormone (PRTH) types. Since similar mutations have been identified in tri-iodothyronine (T3) receptor (TR) beta gene in GRTH and PRTH, and since considerable overlap has been seen in the clinical manifestations in patients with GRTH and PRTH, two subtypes of RTH are now considered to be a continuous spectrum with the same genetic defect. A point mutation at amino acid Arg 338 to Trp (R338W) which we identified in a patient with PRTH is very interesting, since R338W has been found in several other patients with PRTH, raising the possibility that this mutation may tend to associate with a phenotype of PRTH. In our previous study, we found that R338W had relatively less impaired transcriptional potency, weaker dominant negative activity on various T3 response elements and poor homodimer formation, as compared with another GRTH mutant. In this study, to investigate the functional properties of R338W further, especially in terms of the relation between transcriptional activity and dimer formations, we introduced the R338W mutation into the mutant receptors, K443E and F451X, constructing the double mutants, R338W/K443E and R338W/ F451X. Both R338W/K443E and R338W/F451X showed negligible T3 binding and transcriptional activities. The dominant negative activities of K443E and F451X were, however, significantly weakened by introducing the R338W mutation. As a control, a double mutant G345R/K443E was constructed by introducing a point mutation, G345R, located in the same exon 9 as R338W, into the K443E mutant. Dominant negative activity did not differ between G345R/K443E and K443E. Homodimer formation was significantly reduced in the double mutants containing R338W, but not G345R. In summary, introducing the R338W mutation, but not G345R, into the mutant TR significantly weakened the dominant negative activity, despite further impairment of the T3 binding and transcriptional activities.

Cells, Cultured↗