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151.
Saki Itonori Kazuya Hidari Yutaka Sanai Masaru Taniguchi Yoshitaka Nagai 《Glycoconjugate journal》1989,6(4):551-560
The effect of the chain length of the fatty acid residue of the ceramide moiety of ganglioside GM3 on the binding ability of monoclonal antibody M2590, which is specific for the carbohydrate structure of GM3-ganglioside, was examined by means of a direct binding assay on thin layer chromatography plates (TLC immunostaining) and a quantitative enzyme-linked immunosorbent assay (ELISA). Derivatives of GM3 with a long fatty acid chain reacted with the M2590 antibody, but those with a short fatty acid chain showed no reaction in either assay system. These results suggested that the acyl fatty acid moiety of the ganglioside played an important role in the formation or maintenance of the antigenic structure of the carbohydrate moiety of the ganglioside. 相似文献
152.
Cecilia PC Soh Alastair SR Donald James Feeney Walter TJ Morgan Winifred M Watkins 《Glycoconjugate journal》1989,6(3):319-332
The tetrasaccharides GalNAcß1-4[NeuAc2-3]Galß1-4Glc and GalNAcß1-4[NeuAc2-3]Galß1-4GlcNAc were synthesised by enzymic transfer of GalNAc from UDP-GalNAc to 3-sialyllactose (NeuAc2-3Galß1-4Glc) and 3-sialyl-N-acetyllactosamine (NeuAc2-3Galß1-4GlcNAc). The structures of the products were established by methylation and1H-500 MHz NMR spectroscopy. In Sda serological tests the product formed with 3-sialyl-N-acetyllactosamine was highly active whereas that formed with 3-sialyllactose had only weak activity. 相似文献
153.
Linda Owers Narhi Michael F. Rhode Pamela Hunt Tsutomu Arakawa 《Journal of Protein Chemistry》1989,8(5):669-677
The limited proteolysis of human recombinant TNF- by trypsin yields two stable products resulting from cleavage after Arg6 and Arg44. In solution these two products remain associated together in a trimer with a Stokes' radius slightly greater than the radius of intact TNF- and, therefore, could not be separated from each other under nondenaturing conditions. This limited digest retains at least 20% of the activity of the original TNF- sample, and has a tertiary structure that is similar to that of the native protein by circular dichroism. On the other hand, incorrectly folded, inactive TNF- undergoes extensive digestion following similar treatment with trypsin. These results indicate that the active form of TNF- has a tight core structure which is maintained afterN-terminal cleavage and removal. 相似文献
154.
Ossarath Kol Colette Brassart Geneviéve Spik Jean Montreuil Stéphane Bouquelet 《Glycoconjugate journal》1989,6(3):333-348
We have previously shown that an endo--N-acetylglucosaminidase (EC 3.2.1.96) named Endo B, isolated from culture filtrates of the basidiomyceteSporotrichum dimorphosporum cleaves asialo-, and to some extent, monosialylated bi-antennary glycans of theN-acetyllactosamine type linked to the asparagine residue of peptide or protein moieties [Bouquelet S, Strecker G, Montreuil J, Spik G (1980) Biochimie 62:43–49]. In the present paper, the substrate specificity of the enzyme towards oligomannoside and hybrid type glycans has been analyzed. The results obtained indicate that ovalbumin glycopeptides containing four to seven mannose residues and bovine lactotransferrin glycopeptides containing four to nine mannose residues were completely hydrolyzed by the enzyme. The degree of cleavage was variable among hybrid type structures, since glycopeptides containing the following glycans: (Gal)1(GlcNAc)3(Man)5(GlcNAc)2; (GlcNAc)3(Man)5(GlcNAc)2; (GlcNAc)3(Man)4(GlcNAc)2 were not hydrolyzed by the enzyme while the percentage of hydrolysis of a glycopeptide containing (GlcNAc)2(Man)5(GlcNAc)2 glycan reached 90%. The bovine lactotransferrin was partially deglycosylated (40%) in the absence of non-ionic detergent while native ovalbumin glycoprotein was not hydrolyzed by the enzyme.The oligomannoside-and theN-acetyllactosamine-type degrading activities present in the culture filtrates were not separated at any step of the purification procedure. Both activities were eluted as a single component with an apparent molecular mass of 89 kDa suggesting that they are located on the same enzyme molecule.Endo B represents a powerful tool for removing oligomannoside-andN-acetyllactosamine-type glycans fromN-glycopeptides andN-glycoproteins. Moreover, advantages in the use of Endo B in a soluble form as well as in an immobilized form result in its high activity and in its stability to heat denaturation and storage.Abbreviations Gal
d-galactose
- Man
d-mannose
- GlcNAc
N-acetyl-d-glucosamine
- Con A
concanavalin A
- Asn
asparagine
- GLC
gas liquid chromatography
- TLC
thin layer chromatography
- Endo
endo--N-acetylglucosaminidase
- Endo B
endo--N-acetylglucosaminidase isolated fromSporotrichum dimorphosporum
- PBE
polybuffer exchanger
- SDS-PAGE
sodium dodecylsulfate-polyacrylamide gel electrophoresis 相似文献
155.
156.
Belinda Martineau H. Jane Smith Caroline Dean Pamela Dunsmuir John Bedbrook Laurens J. Mets 《Plant molecular biology》1989,13(4):419-426
We report the successful transformation, via Agrobacterium tumefaciens infection, and regeneration of two species of the genus Flaveria: F. brownii and F. palmeri. We document the expression of a C3 plant gene, an abundantly expressed ribulose 1,5-bisphosphate carboxylase/oxygenase small subunit gene isolated from petunia, in these C4 plants. The organ-specific expression of this petunia gene in Flaveria brownii is qualitatively identical to its endogenous pattern of expression. 相似文献
157.
Ulrik Pedersen-Bjergaard Lars B geskov Nielsen Kai Jensen Lars Edvinsson Inger Jansen Jes Olesen 《Peptides》1989,10(6):1147-1152
Neurokinin A (NKA), substance P (SP) and the two peptides combined (SP + NKA) were injected intracutaneously on the forearm and into the temporal muscle of healthy volunteers. Pain intensity, cutaneous wheal and flare responses and tenderness of the temporal muscle were quantitated. SP but not NKA induced cutaneous pain. This relates the algesic effect of SP to the specific N-terminal amino acid sequence of the peptide, not shared by NKA. NKA, however, potentiated the algesic effect of SP as SP + NKA induced a significantly prolonged cutaneous pain sensation. Both peptides induced wheals, but only SP induced flare. These results confirm previous studies relating wheal formation to the identical C-terminal amino acid sequence of the two peptides and flare reaction to the N-terminal part of SP. Injections into the temporal muscle did not cause pain or tenderness. 相似文献
158.
Cellular origin and distribution of transforming growth factor-β1 in the normal rat myocardium 总被引:6,自引:0,他引:6
Dr. M. Eghbali 《Cell and tissue research》1989,256(3):553-558
Summary Transforming growth factor- (TGF-) is a biologically active polypeptide present in normal tissues as well as transformed cells. Two structurally related forms of this peptide are TGF-
1 and TGF-
2. Using freshly isolated cardiomyocytes and non-myocyte heart cells, and a [32P]-labelled cDNA probe to human TGF-
1, we demonstrated that mRNA for TGF-
1 could be detected only in the nonmyocyte fraction of heart cells. In the present study, the distribution of TGF-
1 in the heart was determined by immunofluorescence staining by use of a polyclonal antibody to porcine TGF-
1 in cryostat sections of rat heart. Immunofluorescence staining was intense around the blood vessels and radially diffuse in the surrounding myocardium. 相似文献
159.
Dr. P. Redecker 《Cell and tissue research》1989,257(3):657-660
Summary The Mongolian gerbil (Meriones unguiculatus) contains abundant follicles throughout the intermediate lobe (IL) of the pituitary gland in the adult animal. The mode of follicle formation, the nature of the follicle building cells and the distribution of follicles were investigated in semithin sections of the gerbil IL. The sections were stained conventionally, or immunohistochemically with antibodies directed against -melanocyte stimulating hormone (- MSH). Follicular cells were constantly -MSH-negative, and resembled the marginal cells lining the hypophyseal cleft with regard to their cytological and immunohistochemical properties. Moreover, follicular cells appeared to be derived from strands of marginal cells that regularly invaginated deep into the IL. Both follicular and marginal cells often made up cellular clusters. This process coincided with follicle formation and the generation or transport of the colloidal content found inside follicles and the hypophyseal cleft. Although the non-secretory cells of the IL obviously constituted one major source of pituitary colloid in the gerbil, -MSH-positive secretory cells, which occasionally were found to be discharged into the cleft cavity, might contribute to the colloidal contents. 相似文献
160.
Archaeoglobus fulgidus is an extremely thermophilic archaebacterium that can grow at the expense of lactate oxidation with sulfate to CO2 and H2S. The organism contains coenzyme F420, tetrahydromethanopterin, and methanofuran which are coenzymes previously thought to be unique for methanogenic bacteria. We report here that the bacterium contains methylenetetrahydromethanopterin: F420 oxidoreductase (20 U/mg), methenyltetrahydromethanopterin cyclohydrolase (0.9 U/mg), formyltetrahydromethanopterin: methanofuran formyltransferase (4.4 U/mg), and formylmethanofuran: benzyl viologen oxidoreductase (35 mU/mg). Besides these enzymes carbon monoxide: methyl viologen oxidoreductase (5 U/mg), pyruvate: methyl viologen oxidoreductase (0.7 U/mg), and membranebound lactate: dimethylnaphthoquinone oxidoreductase (0.1 U/mg) were found. 2-Oxoglutarate dehydrogenase, which is a key enzyme of the citric acid cycle, was not detectable. From the enzyme outfit it is concluded that in A. fulgidus lactate is oxidized to CO2 via a modified acetyl-CoA/carbon monoxide dehydrogenase pathway involving C1-intermediates otherwise only used by methanogenic bacteria.Non-standard abbreviations APS
adenosine 5-phosphosulfate
- BV
benzyl viologen
- DCPIP
2,6-dichlorophenolindophenol
- DMN
2,3-dimethyl-1,4-naphthoquinone
- DTT
DL-1,4-dithiothreitol
- H4F
tetrahydrofolate
- H4MPT
tetrahydromethanopterin
- CH2
H4MPT, methylene-H4MPT
- CH
H4MPT, methenyl-H4MPT
- Mes
morpholinoethane sulfonic acid
- MFR
methanofuran
- Mops
morpholinopropane sulfonic acid
- MV
methyl viologen
- Tricine
N-tris(hydroxymethyl)-methylglycine
- U
mol product formed per min 相似文献