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121.
122.
Ding W. Wu John G. Duman Chi-Hing C. Cheng Francis J. Castellino 《Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology》1991,161(3):271-278
Summary Four antifreeze proteins (AFPs) were purified from larvae of the beetle Dendroides canadensis. The AFPs are similar in amino acid compositions, having high contents of hydrophilic amino acids (45–55 mol%) and cysteine (16 mol% Cys). Approximately half of the Cys residues form disulfide bridges, and both the disulfide bridges and free sulfhydryls are essential for activity. The N-terminals of the AFPs are blocked. The pH optimum of the AFPs is 7.8, but major loss of activity occurred only at very high pH (12.0). The detergents SDS and Triton X-100 did not inactivate the AFPs. Circular dichroism spectra indicate the presence of both and secondary structures in the AFPs, in addition to a large random structure component.Abbreviations
AFP
antifreeze protein
-
CD
circular dichroism
-
DTT
dithiothreitol
-
HPLC
high pressure liquid chromatography
-
PAGE
polyacrylamide gel electrophoresis
-
PAS
periodic acid Schiff
-
SDS
sodium dodecyl sulfate
-
TFA
trifluoroacetic acid 相似文献
123.
Ding Wen Wu John G. Duman 《Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology》1991,161(3):279-283
Summary Purified antifreeze proteins (AFPs) from the larvae of the beetle Dendroides canadensis do not produce the high levels of antifreeze activity seen in the hemolymph of overwintering larvae, even when the purified AFPs are assayed at very high concentrations. However, addition of certain proteins or agar (at concentrations sufficiently low that the gel state does not result) to the Dendroides AFP resulted in a 2–3-fold increase in activity. A 70-kDa protein with AFP-activating capabilities was purified from Dendroides larvae. Addition of this endogenous activator protein to a 4 mg·ml-1 solution of AFP increased the activity of the AFPs to values comparable to those of the hemolymph of overwintering larvae. Data derived from a modified immunoblot technique demonstrate that the activators bind to the AFP, or vice versa. Formation of this association must allow the AFP to block ice crystal growth by binding to the surface of potential seed crystals in the normal fashion. However, because the AFP-activator complex is much larger than the AFP alone, the complex probably blocks a greater surface area of the crystal and is thus a more efficient antifreeze.Abbreviations
AFP
antifreeze protein
-
BSA
bovine serum albumine
-
DEAE
diethylaminoethyl
-
Ig
immunoglubolin
-
LPIN
lipoprotein ice nucleator
-
PIN
protein ice nucleator
-
SDS
sodium dodecyl sulfate
-
PAGE
polyacrylamide gel electrophoresis
-
TH
thermal hysteresis 相似文献
124.
M Espinosa-Cantellano A Martínez-Palomo 《Biology of the cell / under the auspices of the European Cell Biology Organization》1991,72(3):189-200
The plasma membrane components of the parasitic protozoan Entamoeba histolytica, the causative agent of human invasive amebiasis, have been biochemically and immunologically characterized during the last decade. In addition, genes coding for certain surface proteins have been cloned. In spite of these advances, a unified characterization of plasma membrane antigenic components of the parasite is still required for badly needed advancements in the design of useful diagnostic, epidemiologic, and immunoprophylactic tools. Here we review current knowledge on this issue and address the problem of the considerable variation in the electrophoretic profiles of plasma membrane proteins obtained by different groups. In addition, the differences in the degree of recognition of reported membrane antigens with human immune sera, and the diverse interpretations concerning the possible functions of the surface molecules characterized are discussed. A comparative analysis of plasma membrane proteins of E histolytica trophozoites using three different isolation methods revealed that it is possible to select for specific membrane proteins, depending on the lysis conditions. In our view, the method of Calderón and Avila preserves more proteins than other methods tested. Using sera from recent cases of invasive amebiasis studied by several laboratories in various geographical areas, a basic antigenic pattern of 11 principal proteins with molecular weights of 220, 170, 150, 125, 97, 80, 60, 45, 20 and 9 kDA was established for the pathogenic E histolytica strain HM1:IMSS, used by most research groups. 相似文献
125.
商陆根中抗真菌蛋白的分离和特性研究 总被引:1,自引:0,他引:1
从美洲商陆(Phytolacca americana L.)夏天采集的2—4年生宿根中分离了二种抗真菌蛋白,称为PAFP-R_1和PAFP-R_2。分离程序包括用盐溶液提取,经CM-Sephadex离子交换层析、凝胶过滤层析和羟基磷灰石柱层析纯化。在PGA培养基上,0.1mg/ml蛋白明显抑制木霉菌丝的生长;但对细菌的增殖,即使1mg/ml也无抑制作用。用SDS-PAGE测得二者的相对分子量各为13kD和15kD,单多肽链,等电点约为5.8。用酚-硫酸法未测出含糖。二种蛋白均高含半胱氨酸(19mol/mol)。用Edman降解法测得二者的N末端均为Ala。秋天采集的根中,这二种蛋白含量均很低,但富含由二条PAFP-R_1肽链以双硫键联结的二聚体,它无抗真菌活性。冬天宿根中抗真菌蛋白主要成分是Mr为17kD的单肽链蛋白。上述蛋白对于人红血球均无凝集活性,因此不是PWM的成分。以上结果说明,商陆根中有多种具抗真菌活性的蛋白,成分随季节发生变化,它们都是不含糖,高含半胱氨酸,分子量小于20kD的单肽链蛋白。 相似文献
126.
127.
Summary Calretinin and calbindin-D28k are two calcium-binding proteins that are present in largely different sets of nerve cells in the central nervous system. Their appearance during development of the chick retina was studied by immunohistochemistry and Western blots. The patterns are mature one day before hatching. Each cell type acquires its characteristic calcium-binding protein several days after its differentiation has started, but in most cases before morphological maturation is complete. There is also an early phase of calbindin immunoreactivity in many immature amacrine cells, and of calretinin immunoreactivity in the presumptive photoreceptor layer, suggesting that these proteins may have distinct functions in differentiating cells.Abbreviations CR+
Immunoreactive for calretinin only
- CB+
immunoreactive for calbindin only
- CR+CB+
immunoreactive for both antisera
- IPL
inner plexiform layer
- OPL
outer plexiform layer 相似文献
128.
Oxidative stress responses were tested in the unicellular cyanobacterium Synechococcus PCC 7942 (R2). Cells were exposed to hydrogen peroxide, cumene hydroperoxide and high light intensities. Activities of ascorbate peroxidase and catalase were correlated with the extent and time-course of oxidative stresses. Ascorbate peroxidase was found to be the major enzyme involved in the removal of hydrogen peroxide under the tested oxidative stresses. Catalase activity was inhibited in cells treated with high H2O2 concentrations, and was not induced under photo-oxidative stress. Regeneration of ascorbate in peroxide-treated cells was found to involve mainly monodehydroascorbate reductase and to a lesser extent dehydroascorbate reductase. The induction of the antioxidative enzymes was dependent on light and was inhibited by chloramphenicol. Peroxide treatment was found to induce the synthesis of eight proteins, four of which were also induced by heat shock.Abbreviations ASC
ascorbate
- DHA
dehydroascorbate
- MDA
monodehydroascorbate
- GSH
reduced glutathione
- GSSG
oxidized glutathione
- ASC Per
ascorbate peroxidase
- DHA red.
dehydroascorbate reductase
- MDA red.
monodehydroascorbate reductase
- GSSG red.
glutathione reductase
- HSP
heat shock proteins
- PSP
peroxide shock proteins
- Cm
chloramphenicol 相似文献
129.
Mario Lachapelle Moise Bendayan 《Virchows Archiv. B, Cell pathology including molecular pathology》1991,60(1):105-111
Actin and alpha-actinin immunoreactive sites have been localized at the electron microscope level by the protein A-gold immunocytochemical
technique in podocytes of normal and nephrotic rat renal tissues. In normal renal glomeruli, fibrillar networks located in
the core of foot processes or bundles of micro filaments interconnecting them were found to be labelled for these two cytoskeletal
proteins. On the other hand, in nephrotic renal glomeruli, concomitant with the loss of podocytic foot processes a reorganization
of the podocytic cytoskeleton and a concentration of some of its elements into thick uniform bands was observed. Actin and
alpha-actinin were revealed in these bands. Control experiments confirmed the specificity of the labelling obtained. Our results
suggest that normal podocytes contain an actin-based contractile system that might contribute to the maintenance of the particular
cell shape of these cells and that the rearrangement of the podocytic cyto-skeleton occurring in the nephrotic syndrome might
account for the changes in the foot processes and contribute to the alteration in glomerular function.
This work was supported by grants from the Medical Research Council of Canada 相似文献
130.
Recombinant Trypanosoma cruzi antigens and Chagas' disease diagnosis: analysis of a workshop 总被引:1,自引:0,他引:1
M.J. Levin J. Franco da Silveira A.C.C. Frasch M.E. Camargo S. Lafon W.M. Degrave R. Rangel-Aldao 《FEMS microbiology letters》1991,89(1):11-20
Abstract A workshop organized by the Ibero-American Project of Biotechnology evaluated the diagnostic potential of several cloned Trypanosoma cruzi recombinant antigens for Chagas' disease serodiagnosis. A set of recombinants, Antigen 2, Antigen 13, SAPA, H49, A13, JL5, JL7, JL8, JL9, and RA1 provided by three different South American laboratories were probed with a panel of 236 South American serum samples. Antigens JL7, H49, Antigen 2, and A13 scored as the best diagnostic recombinant reagents. The results suggested that the main advantage of using cloned peptides for chronic Chagas' disease diagnosis resided in their highly specific immunoreactive properties. 相似文献