Nucleotide sequence of a Brassica napus Clp homolog.
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ATP-Dependent Proteases↗
Biomedical subjects
K Ko
Publications and source records attributed to K Ko.
A method for monitoring intracranial temperature via tunneled ventricular catheter: technical note.
A simple technique for monitoring intracerebral temperature in humans via a ventricular catheter is described. This differs from a previously described method by enabling such measurements to be accomplished with a commercially available thermistor, a standard ventricular catheter, and common hospital supplies. In contrast to the earlier device, this system allows for the subcutaneous tunneling of the distal ventricular catheter. This is an easily assembled and cost-effective technique with which to conduct investigations on human intracerebral temperature.
Brain Injuries↗
Identification of an uncleavable targeting signal in the 70-kilodalton spinach chloroplast outer envelope membrane protein.
A cDNA clone encoding a cognate 70-kDa heat shock protein from the spinach chloroplast outer envelope (SCE70) was recently characterized (Ko, K., Bornemisza, O., Kourtz, L., Ko, Z. W., Plaxton, W. C., and Cashmore, A. R. (1992) J. Biol. Chem. 267, 2986-2993). Initial studies revealed that SCE70 is targeted to the chloroplast outer envelope membrane without further processing. To determine whether SCE70 possesses a "targeting domain," we tested the targeting ability of SCE70 proteins with various carboxyl- and amino-terminal deletions. Carboxyl-terminal deletions of up to 60% of the protein had no apparent effect on the targeting ability of SCE70. Amino-terminal deletions abolished targeting to the chloroplast except when the extreme NH2-terminal 48-amino acid sequence was retained. We further assessed the chloroplast-targeting ability of the NH2-terminal 48 amino acids by fusing to the foreign protein, mouse dihydrofolate reductase (DHFR). The resulting fusion protein, SCE70-DHFR, was localized to the outer envelope membrane of isolated chloroplasts. SCE70-DHFR exhibited targeting characteristics similar to native SCE70. The targeting of SCE70-DHFR was inhibited effectively by anti-SCE70 antibodies. Immunoprecipitation and chemical cross-linking experiments revealed that SCE70-DHFR is targeted to the same complex as SCE70 in the chloroplast envelope. These results suggest that the extreme NH2 terminus of SCE70 is required for directing SCE70 to a destination in the chloroplast outer envelope membrane, possibly through assembling the polypeptide into a protein complex.
Chloroplasts↗
Spontaneous development of anti-collagen type II antibodies with NTA, and anti-DNA antibodies in senescence-accelerated mice.
We previously reported that circulating natural thymocytotoxic autoantibody (NTA) and IgG-anti-DNA antibodies were the major serological characteristics of a substrain of SAM, SAM-P/1. We present here a study of ageing in which we further measured and compared various kinds of circulating IgG antibodies including anti-collagen type II, rheumatoid factor (RF), and anti-2,4,-dinitrophenol (DNP), between SAM-P/1 and control SAM-R/1 mice. The results showed that age-associated increases in anti-collagen type II antibodies in SAM-P/1 were distinctively higher than those in SAM-R/1 when the mice were over 4 months of age, and the increases were significantly correlated with increases in NTA, anti-DNA antibodies, RF activities and anti-DNP antibodies. Anti-collagen type II antibody activity was not significantly inhibited by preincubating the antibodies with DNA, IgG-Fc and DNP-BSA samples. These findings suggest that antibodies specifically directed against collagen type II can be produced in a background of polyclonal B cell activation, and that these antibodies in association with NTA and anti-DNA antibodies may play a pathogenic role in the development of accelerated senescence in SAM-P/1 mice.
Aging↗
Sarcoplasmic reticulum fast CA(2+)-pump and myosin heavy chain expression in extraocular muscles.
PURPOSE: To investigate the distribution of the sarcoplasmic reticulum fast-twitch Ca(2+)-pump (Ca(2+)-adenosine triphosphatase [Ca(2+)-ATPase]) in extraocular muscle fiber populations of the rat and rabbit, using fast Ca(2+)-adenosine triphosphatase-specific monoclonal antibodies. METHODS: Adult female rats and rabbits were killed with overdose of sodium pentobarbital, and the extraocular muscles were rapidly frozen in isopentane cooled with liquid nitrogen. Serial transverse frozen sections were cut and labeled respectively with monoclonal antibodies specific for various isoforms of myosin heavy chain, and monoclonal antibodies specific for the fast isoform of the sarcoplasmic reticulum Ca(2+)-ATPase. RESULTS AND CONCLUSIONS: In rat extraocular muscle, fast type 2 myosin heavy chain and fast Ca(2+)-ATPase are coexpressed in both singly- and multiply-innervated fibers of the orbital layer near the endplate band, but distally both proteins drop out entirely, indicating that the contraction and relaxation properties of orbital fibers vary along their length. Also, fast Ca(2+)-ATPase persists more distally in orbital fibers than does type 2 myosin heavy chain, suggesting that their joint expression is not directly coregulated. This conclusion is reinforced in rabbit, whose orbital multiply-innervated fibers express fast Ca(2+)-ATPase but do not express type 2 myosin heavy chain at all. Fast Ca(2+)-ATPase is expressed in all global layer fast-twitch singly-innervated fibers, but not in global multiply-innervated fibers. In light of known differences in myosin heavy chain expression, the two populations of multiply-innervated fibers must differ with respect to both contractility and relaxation.
Animals↗
Carboxyl-terminal sequences can influence the in vitro import and intraorganellar targeting of chloroplast protein precursors.
The transit peptide of the lumenal 33-kDa oxygen-evolving polypeptide (OEE1) is capable of directing the import and targeting of the foreign protein dihydrofolate reductase (DHFR) to the thylakoid lumen. The import results from the first part of this study indicate that methotrexate cannot block the import or intraorganellar targeting of OEE1-DHFR in chloroplasts in contrast to that reported for the import of cytochrome oxidase subunit IV (COXIV)-DHFR in mitochondria. These results suggest that the fusion of the OEE1 transit sequence to DHFR affected the protein's methotrexate binding properties. We further examined and compared the transport characteristics of a number of carboxyl-terminal truncated native chloroplast precursors to determine whether carboxyl domains contribute to the import and intraorganellar targeting mechanism of these proteins. The plastid precursors chosen for this study are targeted to one of the following chloroplast compartments: the stroma, the thylakoid membrane, and the lumen. In most cases, removal of carboxyl domains had a dramatic effect on one or more stages of the translocation pathway, such as import, processing, and intraorganellar targeting. The effects of carboxyl deletions varied from precursor to precursor and were dependent on the extent of the deletion. These combined results suggest that carboxyl domains in the mature part of the proteins can influence the function of the transit peptide, and as a result play an important role in determining the import and targeting competence of chloroplast precursors.
Chloroplasts↗
Isolation and characterization of a cDNA clone encoding a cognate 70-kDa heat shock protein of the chloroplast envelope.
The translocation of proteins into the endoplasmic reticulum, the mitochondrion, and the chloroplast has recently been shown to involve homologues of the highly conserved 70-kDa heat shock protein (HSP70) family. In this study, we have isolated and sequenced a full-length cDNA clone encoding a cognate 70-kDa heat shock protein of the spinach chloroplast envelope (SCE70). The cDNA insert is 2,535 base pairs long and codes for 653 amino acid residues of a protein with a predicted molecular mass of 71,731 daltons. The deduced amino acid sequence shows a high degree of homology with HSP70 proteins from other organisms. Southern genomic and RNA analyses reveal different hybridization patterns than that observed for a heat-inducible 70-kDa protein gene. The protein synthesized from the SCE70 cDNA insert co-migrates with a 70-kDa polypeptide of the chloroplast envelope following sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Western blot analysis and import studies indicate that SCE70 is associated with the chloroplast outer envelope. The import data suggest that SCE70 is targeted to the envelope membrane via a pathway different from other plastidic precursors but similar to that recently reported for outer envelope proteins SOE1 and OM14.
Amino Acid Sequence↗
Normal growth of transgenic tobacco plants in the absence of cytosolic pyruvate kinase.
The coding sequence of the cytosolic isozyme of potato tuber pyruvate kinase (PK) was attached to the transit peptide of the small subunit of pea ribulose-1,5-bisphosphate carboxylase oxygenase and placed under the control of the cauliflower mosaic virus 35S promoter. This construct was transformed into Nicotiana tabacum. Unexpectedly, two primary transformants were recovered in which PK activity in leaves was greatly reduced. The reduction in PK activity appeared to result from the complete absence of the cytosolic form of the enzyme (PK(c)). In addition, no PK(c) could be detected on western blots of leaf extracts. Metabolite analyses indicated that the levels of phosphoenolpyruvate are substantially higher in PK(c)-deficient leaves than in wild-type leaves, consistent with a block in glycolysis at the step catalyzed by PK. PK(c) deficiency in the leaves does not appear to adversely affect plant growth. Analysis of progeny indicates that PK(c) deficiency is a heritable trait. The leaves of PK(c)-deficient transformants have normal rates of photosynthetic O(2) evolution and respiratory O(2) consumption, indicating that these plants are using alternative pathways to bypass PK.
Journal Article↗
[Effect of botulinum A toxin injection in the management of hemifacial spasm].
The present study was carried out on the effect of botulinum A toxin, made in Japan, in the management of 30 hemifacial spasm patients. In almost all cases, alleviation of hemifacial spasm reached its peak in a few days. Availability was 93.3% and its effect lasted about 4 months. Side effects were observed in 12 patients (40%), but no serious complications were observed.
Adult↗
Isolation and characterization of tomato cDNA and genomic clones encoding the ubiquitin gene ubi3.
We report here the isolation and nucleotide sequence of tomato cDNA and genomic clones encoding a ubiquitin extension protein homologous to the yeast gene ubi3. Sites similar to upstream activating sites commonly found in the promoters of yeast ribosomal genes were observed in the tomato promoter. The tomato ubi3 promoter also contained elements found in the rbcS promoter from pea. The transcription initiation site was determined to occur 66 bp upstream of the initiating Met. RFLP mapping revealed that the gene was located on chromosome 1, 23 cM from marker TG301. A ubi3 gene-specific probe hybridized to a single 800 nt transcript. Expression was reduced in heat-shocked plants and plants kept in the dark. Expression was highest in young leaves and immature green fruit and lowest in mature leaves and petals. We isolated the original cDNA clone using an antibody prepared against chloroplast polypeptides. Immunological studies did not detect ubiquitin or ubiquitin extension proteins in the chloroplast. However, higher-molecular-weight chloroplast proteins were detected with ubiquitin antisera suggesting that ubiquitin conjugates are transported into the chloroplast.
Amino Acid Sequence↗
[Late results of TVSI for Ebstein's anomaly].
For 10 adult cases of Ebstein's anomaly, tricuspid valve supraannular imposition (TVSI) without excision of native tricuspid valve were performed and its late results (6 mo. to 11.5 y, mean 6.8 years) were investigated. All cases showed the severe low output syndrome after the surgery. But the postoperative recovery of the cardiac function was smooth and there was no early death nor A-V block. Both the A-V conduction and the RV subvalvular structures were preserved in TVSI and it contributed to the postoperative good contractility of RV. In two cases, the serious postoperative ventricular tachycardia or ventricular fibrillation were experienced, one case required open cardiac massage and 6 hours long assist circulation in the other. In late stage, one patient died 9 years after the TVSI from the failure of the bioprosthetic mitral valve which had been implanted simultaneously, and the other 9 survivors returned to the class I of NYHA classification. LVDd and DS obtained by echocardiography showed significant enlargement; 29 +/- 4.5 mm and 19 +/- 4.1 mm before operation to 42 +/- 3.1 mm and 29 +/- 5.8 mm in late stage respectively. EF showed the tendency of slight decrease from the preoperative supernormal value to the normal range. RVD showed significant decrease after the surgery (58 +/- 4.2 mm to 37 +/- 4.1 mm). Bioprosthetic valve failure in tricuspid position was not experienced. PAC or PVC were seen quite often and 8 of 9 cases received medication for arrhythmia even in late stage.(ABSTRACT TRUNCATED AT 250 WORDS)
Adolescent↗
Allopurinol and ischemia/reperfusion injury: new use for an old drug?
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Allopurinol↗
Diltiazem reduces the contractility of extraocular muscles in vitro and in vivo.
Diltiazem, a Ca-channel blocker that is used clinically for the treatment of hypertension and cardiac arrhythmias, reduces the contractility of extraocular muscles. Exposure of rabbit extraocular muscle to diltiazem in vitro reduces the sustained tension that is generated by the tonic, multiply innervated fibers, and decreases the baseline, or resting, tension of the muscle. When diltiazem is injected into a selected extraocular muscle in the rabbit in vivo, it causes a temporary weakening of the muscle, which is indicated by a deviation of eye position. These in vivo effects are of short duration, are easily reproducible, and vary with dosage. The results of this study raise the possibility that diltiazem may be used as an alternative to the surgical treatment of strabismus and other oculomotor dysfunctions including blepharospasm.
Adenosine Triphosphatases↗
Nucleotide sequence of the psbA gene encoded by the Vicia faba chloroplast genome.
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Amino Acid Sequence↗
Isolation and characterization of cDNAs from Lycopersicon esculentum and Arabidopsis thaliana encoding the 33 kDa protein of the photosystem II-associated oxygen-evolving complex.
We have identified and isolated cDNA clones of the 33 kDa protein of the oxygen-evolving complex (OEE1) from Lycopersicon esculentum (tomato) and Arabidopsis thaliana and determined their nucleotide sequences. The cDNA clones and antibodies prepared against OEE1 were used as probes to examine the expression of the oee1 gene with respect to regulation by light, organ specificity, and ripening stage of tomato fruit. The steady-state mRNA level is regulated by light, being present in light-grown plants and absent in etiolated seedlings. The oee1 transcripts that accumulate during growth in the light were reduced to non-detectable or low levels by a 3-day dark treatment. The oee1 gene also exhibits differential expression in various organs of the tomato plant. Steady-state mRNA level was highest in immature leaves and absent in roots while the protein was most abundant in mature leaves and absent in roots. During tomato fruit ripening oee1 mRNA decreases to a low level in the pericarp while the protein level decreases below detection. The expression of oee1 appears to be under the control of a complex mechanism regulating both the amount of RNA and its translation.
Amino Acid Sequence↗
An immunomodulating protein, Ling Zhi-8 (LZ-8) prevents insulitis in non-obese diabetic mice.
Ling Zhi-8 (LZ-8), a novel and recently discovered immunomodulatory protein having in vivo immuno-suppressive activity, was tested for in vivo effect against Type 1 (insulin-dependent) diabetes mellitus in the nonobese diabetic mouse, the disease having immunologically mediated aetiology in this animal. LZ-8 had mitogenic activity in vitro towards spleen cells of the non-obese diabetic mice as previously shown towards those of DBA/2 mice. Intraperitoneal administration of LZ-8 twice weekly into the mice (10.3-12.6 mg/kg body weight) from 4 weeks of age prevented insulitis and an almost normal number of insulin producing cells were observed. Extreme insulitis and reduction of the number of insulin producing cells were observed in the pancreata of the untreated non-obese diabetic mouse. No cumulative incidence of diabetes mellitus was observed in the LZ-8 treated group, while cumulative incidences of 70% and 60% were observed in an untreated group followed up to 42 weeks of age when the incidence of diabetes was defined as a plasma glucose level of greater than 11 mmol/l and as a urine glucose level of greater than 2+, respectively. T cell subset population analysis was performed to further investigate the action of LZ-8 on the non-obese diabetic mouse which revealed that LZ-8 treatment increased in L3T4'/Lyt-2+ ratio.
Adjuvants, Immunologic↗
Systematic variation in myosin expression along extraocular muscle fibres of the adult rat.
Monoclonal antibodies (McAB) specific for fast (C14) and slow (S58) myosin, and a myosin antigenically similar to neonatal/embryonic myosin in mammals (ALD180), were used to characterize the myosin distribution in orbital layer fibres of rat extraocular muscles (EOM) in relation to innervation patterns. The orbital layer is composed of both singly-innervated (SIF) and multiply-innervated (MIF) fibres. The SIFs have the characteristics of twitch fibres, while the MIFs, in addition to possessing many small endings characteristic of tonic fibres, also have an en-plaque-like innervation in the endplate band resembling that of the adjacent SIFs. Myosin expression in MIFs and SIFs is unusual and varies systematically along the length of the fibres. Both SIFs and MIFs label with ALD180, but this labelling is absent in both fibre types in the endplate band region, where all fibres label with C14. Distally and also proximally to the endplate band, SIFs label with both ALD180 and C14, while the MIFs, innervated by many small, superficial endings in these regions, label with ALD180 only. This pattern of myosin expression could also be demonstrated in isolated fibres. The results are discussed in relation to the hypothesis that both populations of orbital layer fibres express constitutively both fast and the neonatal-like myosin, and that superimposed on this constitutive expression twitch or tonic innervation acts locally to selectively suppress either neonatal-like or fast myosin, respectively.
Adenosine Triphosphatases↗
Complete amino acid sequence of an immunomodulatory protein, ling zhi-8 (LZ-8). An immunomodulator from a fungus, Ganoderma lucidium, having similarity to immunoglobulin variable regions.
The complete amino acid sequence of a novel immunomodulatory protein, ling zhi-8 (LZ-8), isolated from a fungus, Ganoderma lucidium (Kino, K., Yamashita, A., Yamaoka, K., Watanabe, J., Tanaka, S., Ko, K., Shimizu, K., and Tsunoo, H. (1989) J. Biol. Chem. 264, 472-478), was determined by protein sequencing. The polypeptide consists of 110 amino acid residues with an acetylated amino end and has a molecular mass of 12,420 Da including an amino-end blocking group. There is no attachment site for an Asn-linked oligosaccharide chain, consistent with the very low carbohydrate content of LZ-8. These results indicate that the native form of LZ-8 with a molecular mass of 24 kDa is a homodimer of the LZ-8 polypeptide whose sequence is described here. Furthermore, the LZ-8 chain shows considerable similarity to the variable region of immunoglobulin heavy chain both in its sequence and in its predicted secondary structure. The interesting possibility that LZ-8 is related to an ancestral protein of the immunoglobulin superfamily is also discussed.
Adjuvants, Immunologic↗