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1.
The specificity of CTP:phosphocholine cytidylyltransferase from rat liver for phosphorylated bases has been investigated. The apparent Km for phosphocholine was 0.17 mM. As the number of methyl substituents on the phospho-base decreased, the apparent Km increased: 4.0 mM for phosphodimethylethanolamine, 6.9 for phosphomonomethylethanolamine and 68.4 for phosphoethanolamine. The Vmax for the reaction was similar for phosphocholine (12.6 mumol/min per mg protein), phosphomonomethylethanolamine (13.5 mumol/min per mg protein) and phosphoethanolamine (9.2 mumol/min per mg protein). When phosphodimethylethanolamine was the substrate, the Vmax was 3-fold higher (40.3 mumol/min per mg protein). Phosphoethanolamine, phosphomonomethylethanolamine and phosphodimethylethanolamine were competitive inhibitors of the cytidylyltransferase when phosphocholine was used as substrate with Ki values of 18.5 mM, 9.3 mM and 1.5 mM, respectively. The results show that the cytidylyltransferase is highly specific for phosphocholine. 相似文献
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4.
Ravigadevi Sambanthamurthi Siti Hasnah Parman Mohd. Roslan Md. Noor 《Plant Cell, Tissue and Organ Culture》1996,46(1):35-41
Procedures are deseribed for the efficient isolation of protoplasts from a variety of oil palm (Elaeis guineensis Jacq.) tissues. Various factors including donor source, composition of enzyme mixture and culture medium affected the yield and viability of the protoplasts Polyembryogenic cultures of oil palm were the most suitable starting material in terms of yield, viability and metabolic competence. Pectolyase Y-23 in association with cellulase and hemicellulase was required for the efficient release of protoplasts from the oil palm tissues. Limited cell division to form microcallus was observed at very low frequency (<0.01%) when glutathione and catalase were incorporated in the culture medium.Abbreviations 2,4-d
dichlorophenoxyacetic acid
- DTT
dithiothreitol
- MES
2[N-morpholino] ethanesulphonic acid
- NAA
1-naphthalene acetic acid
- PVP
polyvinylpyrrolidone 相似文献
5.
Summary The occurrence of cells resembling mammalian Langerhans cells in the avian epidermis was studied by ATPase histochemistry, Ia immunoreactivity and electron microscopy. The existence of MHC class II antigen-(Ia) expressing, ATPase-positive dendritic cells, which are ultrastructurally similar to mammalian Langerhans cells except for the absence of Birbeck granules, was demonstrated. These cells may be a basic component of the immune system of birds. 相似文献
6.
M Anissimova L Yaouancq F Noor M-A Badet-Denisot B Badet 《The journal of peptide research》2003,62(2):88-95
A direct continuous UV-Vis spectrophotometric assay has been developed for VanX, a D-alanyl-D-alanine aminodipeptidase necessary for vancomycin resistance. This method is based on the hydrolysis of the alternative substrate D-alanyl-alpha-(R)-phenylthio-glycine D-Ala-D-Gly(S-Ph)-OH (H-DAla-DPsg-OH, 5a). Spontaneous decomposition of the released phenylthioglycine generates thiophenol, which is quantified using Ellman's reagent. The dipeptide behaved as an excellent substrate of VanX, exhibiting Michaelis-Menten kinetics with a kcat of 76 +/- 5/s and a KM of 0.83 +/- 0.08 mm (kcat = 46 +/- 3/s and KM = 0.11 +/- 0.01 mm for D-Ala-D-Ala). Determination of the kinetic parameters of the previously reported mechanism-based inhibitor D-Ala-D-Gly(SPhip-CHF2)-OH (H-D-Ala-DPfg-OH, 5c) [Araoz, R., Anhalt, E., René, L., Badet-Denisot, M.-A., Courvalin, P. & Badet, B. (2000) Biochemistry 39, 15971-15979] using the substrate reported in the present study yielded values of Kirr of 22 +/- 1 microM and kinact of 9.3 +/- 0.4/min in good agreement with values previously obtained in our laboratory (Kirr = 30 +/- 1 mm; kinact = 7.3 +/- 0.3/min). In addition, inhibition by the competing substrate D-Ala-D-Ala resulted in determination of a Ki = 70 +/- 6 microM close to the previously reported KM value. These results demonstrate that the present assay is a convenient, rapid and sensitive tool in the search for VanX inhibitors. 相似文献
7.
In this paper we describe a fast and mild method based on the use of a unique cation exchanger and buffers containing ethylene glycol and salt for the purification of the myelin basic protein (MBP; MW 18.5 kDa). MBP thus purified hydrolyses catalytically p-nitrophenyl acetate. This esterase activity facilitates not only the purification of MBP but also indicates that probably it is in its native state, i.e. there is a good chance that the purified molecules are structurally and chemically identical. This is a prerequisite to obtain crystals appropriate for x-ray diffraction and other studies.Abbreviations used MBP
myelin basic protein
- MW
molecular weight
- kDa
kilo Dalton
- octyl-POE
n-octylpolydisperse oligooxyethylene
- CHAPS
3-3-cholamidopropyl dimethylammonio-1-propane-sulfonate
- CTAB
cetyltrimethylammonium bromide
- SDS
sodium dodecyl sulfate
- SDS-PAGE
polyacrylamide gel electrophoresis in the presence of SDS
- G 3707
heptaoxyethylene lauryl ether
- TWEEN-20
polyoxyethylenesorbitan-monolaurat
- EDTA
ethylenediaminetetraacetic acid
- HEPES
N-(2-hydroxyethyl)-piperazine-N-(2-ethanesulfonic acid) 相似文献
8.
In model experiments with Bonus barley,where the growth of a fraction of the seedlings is retarded by a raised thiol content, obtained through treatment with 2-mercaptoethanol, stimulation of growth is reproducibly induced by low doses of γ-rays. As illustrated by a probit representation of seedling heights, and by the decrease of coefficients of variation, the radiostimulation is restricted to that fraction of seedlings which exhibits a suppressd growth. The validity of the model to natural situations was sustained in studies of the variety Impala, which was shown by Fendrik and Bors to respond regularly to stimulating doses of X-rays. In this variety the response was shown to be restricted to a sub-population of seedlings with retarded growth, i.e. the same pattern as observed in thiol-treated Bonus barley. 相似文献
9.
The effect of salinity on reproductive physiology of wheat wasinvestigated. One set of wheat plants was subjected to increasingsalt levels up to a certain concentration, whereas another setwas given the same concentration in a single application. Theformer was called "gradual" and latter "shock" treatment. Theireffects on pollen viability, germination and activity of starchsynthetase were studied. Gradual treatment seemed to reducethe toxic effects of salts on the viability of pollen grainsand their germination. Seeds obtained from the two sets weregerminated in the same salinities in which their plants hadbeen growing, and the results were compared with those of seedsobtained from control plants growing in a non-saline medium.The seeds of plants from the gradual treatment were better suitedfor germination on a saline medium than those from the shocktreatment or the control group. Salt treatment also increasedthe activity of starch synthetase at the midmilky stage in developinggrains. This phenomenon was considered essential for synthesisof starch in a saline environment. The increase in Na+ and Cl and decrease in K+ contentsof wheat grains in both treatments suggest that the effect ofsalinity on the physiological phenomenon studied is due to changesin the ionic content of the plants.
1 In partial fulfilment of a Ph.D. degree from the Universityof Karachi, Pakistan.
2 Professor of Botany, Director of Research Projects, Head,Plant Physiology Section, University of Karachi, Pakistan. (Received July 11, 1977; ) 相似文献
10.
On a new charophyte from India 总被引:1,自引:1,他引:0
Chara indica is described as a new species on morphological and cytological grounds.Part of Ph. D. Thesis of Ranchi University. 相似文献