Purification and characterization of a base non-specific and adenylic acid preferring ribonuclease from the fruit bodies of Lentinus edodes.
Explore the source record for details and available documents.
Adenosine Monophosphate↗
Biomedical subjects
M Irie
Publications and source records attributed to M Irie.
Purification and properties of a novel surface-active agent- and alkaline-resistant protease from Bacillus sp. Y.
In the course of a search for an alkaline stable protease for industrial use, an alkaline protease (protease BYA) was isolated from an alkalophilic Bacillus sp. Y, and its properties were characterized. Its optimum pH was pH 10.0-12.5, when casein was used as a substrate. In addition to the stability of protease BYA at pH 6.5-13.0, it was also very stable towards various surface-active agents, such as sodium dodecyl sulfate and sodium linear alkylbenzene sulfonate. Protease BYA was most active at 70 degrees C. The isoelectric point (pI) of protease BYA was about 10.1. Protease BYA was characterized as a serine protease because of its sensitivity to phenylmethanesulfonyl fluoride and diisopropyl fluorophosphate. The protease seems to be related to proteases of the subtilisin family, such as subtilisin BPN', subtilisin Carlsberg, and No. 221 protease.
Amino Acid Sequence↗
Primary structure of a nuclease (nuclease PA3) from a Penicillium sp.
The complete primary structure of a nuclease from a Penicillium sp. [nuclease PA3 (Kazama et al., Chem. Pharm. Bull., 38, 3081 (1990)] was determined. The sequencing was done by analysis of the peptides generated by digestion of reduced and carboxymethylated nuclease PA3 (RCM nuclease PA3) with lysylendopeptidase, and by digestion with staphylococcal V8 protease or chemical cleavage with BrCN. It consisted of 270 amino acid residues and carbohydrate moieties attached to the 92nd, 138th, 184th, and 197th asparagine residues. The molecular weight of the protein moiety deduced from the sequence was 29,211. It contains four half cystine residues. The amino acid sequence was identical with that of P1 nuclease from Penicillium citrinum [K. Maekawa, S. Tsunasawa, G. Dibo, and F. Sakiyama, Abstracts of Papers, the 62nd Meeting of the Biochemical Society of Japan, Seikagaku, 61, 1013 (1989)] except that the 190th Thr residue was Ile in P1 nuclease.
Amino Acid Sequence↗
[Psychosomatic study of inhalation therapy in patients with bronchial asthma. I. Evaluation by questionnaire].
We investigated the use of metered-dose beta agonist inhalers (MDIs) by means of 17-item questionnaires from the standpoint of psychosomatic medicine in 56 subjects with bronchial asthma. The results can be summarized as follows: 1) Many asthmatics with neurotic characters used MDIs frequently; Thirteen percent of them suffered adverse effects. The number of times the inhalers were used increased in proportion to the severity and the duration of illness. Fifteen percent of the subjects used MDIs frequently because of anxiety. 2) Fifty percent of the subjects, most of them had neurotic or alexithymic characters, used MDIs without suffering from dyspnea or wheezes; the subjects who used MDIs because of anxiety increased in proportion to the duration of illness. 3) Seventy eight percent of the subjects were anxious about asthmatic attack when they did not carry any MDI; sixty two percent of the subjects, most of them had neurotic or alexithymic characters, actually experienced asthmatic attack. 4) Seventy three percent of the subjects used MDIs secretly; sixty percent of the subjects did not like to inhale in the presence of others. 5) There were some subjects who acted incorrectly on or after asthmatic attack when MDIs were not effective.
Adolescent↗
[Psychosomatic study of inhalation therapy in patients with bronchial asthma. II. Relationship between subjective self assessment of dyspnea and peak expiratory flow rates].
The degree of dyspnea when using metered-dose inhalers (MDIs), and the relationship between subjective self assessments and objective peak expiratory flow rates were examined by means of visual analogue scales and mini wright peak flow meters in 16 subjects with asthma. Three subjects who were neurotic in character were found to have a poor perception of the degree of airway obstruction after inhalation compared with objective changes in their condition. Two subjects, one neurotic and one alexithymic, assessed the degree of dyspnea to be worse after inhalation, when objectively there were few changes. However, one neurotic subject perceived an improvement in his condition when, in fact, there was little change. Furthermore, some severe cases used MDIs in the rogressive state of airway obstruction. In conclusion, we must pay attention to patients' characters when determining treatment in order to prevent overuse of MDIs or delays in treatments including the use of MDIs.
Adult↗
Long-term follow-up investigation of the effects of the biopsychosocial approach (BPSA) to bronchial asthma.
The recent rapid increase in the number of allergic patients is becoming a social problem. Studies of the causes of this phenomenon involve various fields, with much attention focused on finding new antigens in food, air, articles encountered in daily living, etc. Recent studies of psychoneuroimmunology (PNI) also suggest a strong influence of emotions on allergic reactions. The number of allergic patients is increasing in all civilized countries without exception, and the stress prevalent in modern civilized society is related to this increase. Modern allergology does not yet have sufficient countermeasures for such stress states. We applied a biopsychosocial approach (BPSA) to treatment programs for allergic disease, incorporating treatment of physical and psychosocial problems en bloc. We studied the long-term effects of BPSA therapy on 82 patients who were treated for more than 3 months in the hospital and were examined 2 to 3 years after discharge. Results showed that more than 80% of patients maintained improvement and 45% of those with intractable asthma were able to withdraw from steroid hormones. BPSA achieved better results than those with standard medication administered only to the body. Improvements after treatment included physical changes, normalization of MV (microvibration) type, decreased levels of plasma histamine, and normal circadian rhythms of lymphocyte subsets. These changes reflect part of the physical mechanisms by which BPSA improves asthma symptoms. From a psychological view point, the patients' feelings, personal relations, behavior, etc. were changed after BPSA, allowing a new life style and improved QOL. It is important for asthma patients to maintain good overall condition over long periods. After BPSA, 80% of our patients were able to do so. It is difficult for the therapist to approach asthma from different aspects at once, including biological, psychological, and social, so we developed a five-stage program of BPSA therapy and found that this obtained favorable results.
Adult↗
Serum profiles of steroid hormones in patients with Cushing's syndrome determined by a new HPLC/RIA method.
We developed a method for simultaneously measuring steroid hormones in very small volumes of serum, using a combination of high-performance liquid chromatography (HPLC) and radioimmunoassay (RIA). By this method, aldosterone, cortisol, 11-deoxycortisol, estrone, estradiol, androstenedione, dehydroepiandrosterone, deoxycorticosterone, 17-hydroxyprogesterone, testosterone, pregnenolone, and progesterone could be determined in a single 100-microL aliquot of serum from normal adults and patients with Cushing's syndrome. The steroid profile associated with Cushing's syndrome caused by adrenal adenoma was quite distinct from that associated with the syndrome caused by adrenal hyperplasia. Serum concentrations of androstenedione, dehydroepiandrosterone, estrone, estradiol, 17-hydroxyprogesterone, pregnenolone, and testosterone were significantly higher in patients with adrenal hyperplasia than in those with an adenoma. We compared the results of this HPLC/RIA method with those of 125I RIAs. The use of a HPLC/RIA system to obtain an accurate and sensitive profile of a range of serum steroids, as described here, obviates the need for large volumes of blood.
Chromatography, High Pressure Liquid↗
[A case of Ménétrier's disease complicated during the clinical course of bronchial asthma].
Explore the source record for details and available documents.
Adult↗
[Evaluation of myocardial blood flow and cardiac functional change in acromegaly].
We studied 201Tl myocardial scintigraphy and 99mTc-HSA gated blood pool scan to evaluate the influence of growth hormone on myocardial blood flow and hemodynamics in 16 patients with acromegalies. Contraction properties were almost within normal limit, but cardiac output was in high output state, because left ventricular ejection fraction (LVEF), right ventricular ejection fraction (RVEF) and cardiac output were 47.8 +/- 17.1%, 37.3 +/- 14.9%, and 6.4 +/- 1.7 L/min, respectively. Otherwise, 201Tl myocardial images revealed 62.5% no redistributed hypoperfusion, 18.8% redistributed hypoperfusion, 56.3% myocardial hypertrophy and 25.0% left ventricular enlargement. These data suggested the high incidence of acromegalic cardiomyopathy due to primary or secondary growth hormone effect.
Acromegaly↗
The structure of the asparagine-linked sugar chains of bovine brain ribonuclease.
The asparagine-linked sugar chains of bovine brain ribonuclease were quantitatively released as oligosaccharides from the polypeptide backbone by hydrazinolysis. After N-acetylation, they were converted into radioactively-labeled oligosaccharides by NaB3H4 reduction. The radioactive oligosaccharide mixture was fractionated by ion-exchange chromatography, and the acidic oligosaccharides were converted into neutral oligosaccharides by sialidase digestion. The neutral oligosaccharides were then fractionated by Bio-Gel P-4 column chromatography. Structural studies of each oligosaccharide by sequential exoglycosidase digestion in combination with methylation analysis revealed that bovine brain ribonuclease showed extensive heterogeneity. It contains bi- and tri-antennary, complex-type oligosaccharides having alpha-D-Manp-(1----3)-[alpha-D-Manp-(1----6)]-beta-D-Manp -(1----4)-beta-D- GlcpNAc-(1----4)-[alpha-L-Fucp-(1----6)]-D-GlcNAc as their common core. Four different outside oligosaccharide chains, i.e., beta-D-Galp-(1----4)-beta-D-GlcpNAc-(1----, alpha-Neu5Ac-(2----6)-beta-D- Galp-(1----4)-beta-D-GlcpNAc-(1----, alpha-Neu5Ac-(2----3)-beta-D-Galp-(1----4)- beta-D-GlcpNAc-(1----, and alpha-D-Galp-(1----3)-beta-D-Galp-(1----4)-beta-D-GlcpNAc-(1----, were found. The preferential distribution of the alpha-D-Galp-(1----3)-beta-D-Galp-(1----4)-beta-D-GlcpNAc group on the alpha-D-Manp-(1----6) arm is a characteristic feature of the sugar chains of this enzyme.
Animals↗
Adrenalectomy-induced alterations in glucagon binding and lipolysis in isolated rat adipocytes.
Adipocytes from adrenalectomized rats nearly lost their lipolytic response to glucagon concomitant with a 90% decrease in the number of glucagon receptors per cell. Quantitative analysis of the relation between amount of cell-bound glucagon and hormone-stimulated lipolysis revealed that the ability of the remaining 10% of glucagon receptors to induce lipolysis was not impaired. Binding of the beta-adrenergic antagonist [3H]dihydroalprenolol and maximal lipolysis induced by (-)-isoproterenol, (Bu)2cAMP, 3-isobutyl-1-methylxanthine, and adenosine deaminase were reduced only 10 to 20% after adrenalectomy. Furthermore, glucagon-stimulated cAMP production was greatly decreased in adrenalectomized animals, but isoproterenol-stimulated cAMP production was not. Hydrocortisone replacement in adrenalectomized rats only partially prevented the loss of glucagon receptors and glucagon effects on both cAMP production and lipolysis. These findings suggest that lipolytic cascade distal to hormone receptors was not greatly impaired in adipocytes after adrenalectomy and that the unresponsiveness of these cells to glucagon was mostly due to a marked reduction in the number of glucagon receptors.
1-Methyl-3-isobutylxanthine↗
Primary structure of an alkaline ribonuclease from bovine liver.
A pyrimidine base specific and most basic alkaline RNase named RNase BL4 was isolated from bovine liver as a protein showing a single band on slab gel-electrophoresis. The enzyme is most active at pH 7.5. The enzyme was immunologically distinguishable from the known bovine RNases such as pancreatic RNase (RNase A), seminal RNase, kidney non-secretory RNase (RNase K2), and brain RNase (RNase BRb). The primary structure of this pyrimidine base-specific RNase was determined to be less than EDRMYQRFLRQHVDPDETG- GNDSYCNLMMQRRKMTSHQCKRFNTFIHEDLWNIRSICSTTNIQCKNGQMNCHEGVVRV- TDCRETGSSRAPNCRYRAKASTRRVVIACEGNPEVPVHFDK. It consists of 119 amino acid residues, and is 5 amino acid residues shorter than RNase A. The sequence homology of RNase BL4 with RNase A is 46.2%, and optimal alignment of RNase A and RNase BL4 requires five deletions, one at the 24th position, two at the 75th and 76th positions, and two at the C-terminus in RNase BL4. The RNase BL4 was highly homologous with a porcine liver RNase (RNase PL3, 94.1% homology) studied by Hofsteenge et al. (personal communication from Hofsteenge, J., Matthies, R., and Stones, S.R.).
Amino Acid Sequence↗
Antigenic determinants on rat metallothionein: fine epitope mapping for a murine monoclonal antibody and rabbit polyclonal antisera.
In order to determine the epitope of metallothionein (MT) to a murine monoclonal antibody (MT 189-14-7) which had been produced by immunization with rat MT 2 (Kikuchi et al. (1988) Mol. Immunol. 25, 1033-1036), various lengths of synthetic oligopeptides were tested for their inhibitory activities in competitive radioimmunoassay (RIA). The amino-terminal acetylated pentapeptide, AcMDPNC, exhibited an inhibitory activity comparable to that of native MTs, whereas the acetylated tetrapeptide, AcMDPN, and the deacetylated heptapeptide, MDPNCSC, were much less inhibitory. The results suggest that the major part of the epitope structure of MT to the MT 189-14-7 monoclonal antibody is located within the amino-terminal acetylated pentapeptide, AcMDPNC. The specificities of polyclonal rabbit anti-MT antisera raised against the same immunogen were also determined by using various animal MTs and synthetic peptides as inhibitors in the RIA. Among three antisera tested, two reacted with several amino-terminal oligopeptides similarly to the MT 189-14-7 antibody. The major epitope structures to these polyclonal antibodies were shown to be located within the acetylated tetrapeptide, AcMDPN. Another antiserum contained at least two different populations of antibodies: one consisted of antibodies reactive with the amino-terminal synthetic peptides, while the other was not reactive with them. These results suggest that, in the rabbit also, the amino-terminal region common to various animal MTs can be an epitope to antibodies raised against rat MT, as shown in the mouse. Moreover, the results indicate that the synthetic amino-terminal peptides are useful for determination of the specificity of polyclonal rabbit anti-MT antibodies, which have been widely used for the quantification of MTs.
Amino Acid Sequence↗
Primary structure of a base non-specific and adenylic acid preferential ribonuclease from Aspergillus saitoi.
The complete primary structure of a base non-specific and adenylic acid preferential RNase (RNase M) from Aspergillus saitoi was determined. The sequence was determined by analysis of the peptides generated by digestion of heat-denatured RNase M with lysylendopeptidase, and the peptides generated from RCM RNase M by digestion with staphylococcal V8 protease or chemical cleavage with BrCN. It consisted of 238 amino acid residues and carbohydrate moiety attached to the 74th asparagine residue. The molecular weight of the protein moiety deduced from the sequence was 26,596. The locations of 10 half cystine residues are almost superimposable on those of RNase Rh from Rhizopus niveus and RNase T2 from Aspergillus oryzae which have similar base specificity. The homology between RNase M and RNase Rh and RNase T2 amounted to 97 and 160 amino acid residues, respectively. The amino acid sequences conserved in the three RNases are concentrated around the three histidine residues, which are supposed to form part of the active sites of these RNases.
Adenosine Monophosphate↗
Glucagon inhibits insulin activation of glucose transport in rat adipocytes mainly through a postbinding process.
Incubation of rat adipocytes with 1 microM glucagon plus adenosine deaminase (5 micrograms/ml) inhibited maximally insulin-stimulated 3-O-methyl-D-glucose (MeGlc) transport by approximately 70%, concomitant with 30% and 55% decreases in insulin binding and cellular ATP, respectively. In contrast, under conditions where cellular ATP levels are well preserved (i.e. high albumin concentration in the medium), the inhibition of transport was reduced to about 30%, but that of insulin binding was not. Because depletion of the cellular ATP level by more than 60% by metabolic inhibitors induced 40% or more inhibition of insulin-stimulated MeGlc transport, the greater inhibition of the transport with the low albumin concentration appears to be caused in part by the secondary effect of ATP loss. The relationship between the amount of cell-bound insulin and hormone-stimulated transport activity showed that glucagon does not modulate insulin action at the step of insulin binding to its receptors. Furthermore, glucagon suppressed insulin-stimulated MeGlc transport, mainly through an attenuation of the hormone-induced increase in maximum velocity. The data show that glucagon modulates the process of signal transduction of insulin action. However, the possibility that glucagon directly modulates the process of translocation or the intrinsic activity of the glucose transporters cannot be eliminated.
3-O-Methylglucose↗
Epidermal growth factor (EGF) receptors in human chorionic gonadotropin-producing tumor: transplantation in nude mice and the effect of EGF on tumor growth.
We examined the presence and characteristics of epidermal growth factor (EGF) receptors in hCG producing tumors (CC-2-JCK) transplanted in female nude mice. We also examined the in vivo effects of EGF on tumor growth. Specific receptors with apparent dissociation constants of 3.89 x 10(-10) and 1.0 x 10(-9) M and binding capacities of 5.96 x 10(-10) and 1.52 x 10(-9) M/mg protein for EGF have been identified in the hCG-producing tumor. [125I]EGF binding to the tumor tissues was time, temperature, and tissue weight dependent and specific. EGF and transforming growth factor-alpha (TGF alpha) competed for [125I]EGF binding, with 50% of the bound [125I]EGF displaced by approximately 0.52 nM EGF and 3.10 nM TGF alpha. TGF beta competed for [125I]EGF binding slightly. Five micrograms of EGF caused an increase in the rate of tumor growth, while 50 micrograms EGF strongly inhibited tumor growth. The concentration of [125I]EGF binding in the tumor treated with low doses of EGF was high, and that in the tumor treated with high doses of EGF was low. These changes in EGF binding were attributable to the changes in the number of high affinity EGF receptors with no significant alteration in binding affinity. In conclusion, the existence of high concentrations of EGF receptors with high affinity and specificity to EGF was demonstrated in an hCG-producing tumor transplanted in nude mice and appeared to be correlated with tumor growth.
Animals↗
Purification and characterization of a nuclease (3'-nucleotidase) from a Penicillium sp.
A nuclease (3'-nucleotidase) similar to P1 nuclease from Penicillium citrinum was purified from a commercial digestive from a Penicillium sp. The activity of the nuclease (PA) was separated to three fractions by diethylaminoethyl-Toyopearl 650M column chromatography, in total yield of 10%. The apparent molecular weight of these three nucleases, PA1, PA2 and PA3 was 35000, 33000, and 32000, respectively. All of them were homogeneous so far as checked by sodium dodecyl sulfate slab gel electrophoresis. The three nucleases differed in carbohydrate content, but their amino acid composition was practically the same, and very similar to that of P1 nuclease. The molecular weight of nuclease PA3, the major component of nuclease PA, was approximately 27000 after digestion by endoglycosidase F. The N-terminal and C-terminal amino acid sequences of nuclease PA3 were determined by Edman degradation and carboxypeptidase(s) digestion, respectively. The nuclease PA3 was inactivated in the presence of 10 mM ethylenediamine tetraacetic acid (EDTA) and 65% of its native enzyme activity restored by the addition of 20 mM ZnCl2. The pH-dependent photooxidative inactivation of nuclease PA3 was accelerated by removal of Zn ion by EDTA or trishydroxymethyl aminomethane, indicating the possible chelation of Zn2+ with some histidine residues.
Amino Acid Sequence↗
Maturation of feedback control of thyrotropin in premature infants.
Serum thyrotropin (TSH), free T4 and free T3 concentrations were measured longitudinally in 26 preterm infants for 14 weeks after birth, using highly sensitive immunoradiometric assays. Serum TSH values on days 4-5 were positively correlated with gestational age and birth weight. In the premature infants of 25 weeks mean gestation, the mean TSH concentrations increased from a very low value of 0.84 microU/ml at 5 days to a peak value of 6.1 microU/ml by 5 weeks of age, then slightly decreased and remained stable. Serum free T4 and free T3 concentrations increased in parallel and free T3 level reached the range of term infants by 6 weeks. Serum free T4/TSH and free T3/TSH ratios began to increase at the 6th week of age. The results suggest that: (i) the thyroid hormone feedback control of pituitary TSH release in the extremely premature infants begins to mature after 6 weeks of postnatal age, (ii) the maturation pattern of the hypothalamic-pituitary-thyroid system in premature infants is similar to that of the intrauterine fetus.
Birth Weight↗