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221.
Alexander A. Kortt Robin E. Guthrie Mark G. Hinds Barbara E. Power Neva Ivancic J. Bruce Caldwell L. Clem Gruen Raymond S. Norton Peter J. Hudson 《Journal of Protein Chemistry》1995,14(3):167-178
The VH domain of anti-influenza neuraminidase antibody NC41, with and without a C-terminal hydrophilic marker peptide (FLAGTM), has been expressed in high yield (15–27 mg/L) inEscherichia coli. Both forms were secreted into the periplasm where they formed insoluble aggregates which were solubilized quantitatively with 2 M guanidine hydrochloride and purified to homogeneity by ion-exchange chromatography. The VH-FLAG was composed of three isoforms (pI values of 4.6, 4.9, and 5.3) and the VH molecule was composed of two isoforms with pI values of 5.1 and 6.7; the difference between the VH isoforms was shown to be due to cyclization of the N-terminal glutamine residue in the pI 5.1 isoform. At 20°C and concentrations of 5–10mg/ml the VH domain dimerized in solution and then partly precipitated, resulting in the broadening of resonances in its1H NMR spectrum. Reagents such as CHAPS,n-octylglucoside, and ethylene glycol, which presumably mask the exposed hydrophobic interface of the VH molecule, prevented dimerization of the VH and permitted good-quality NMR spectra on isotope-labeled protein to be obtained. 相似文献
222.
223.
Alexander A. Bachmanov Danielle R. Reed Yuro Ninomiya Masashi Inoue Michael G. Tordoff R. Arlen Price Gary K. Beauchamp 《Mammalian genome》1997,8(8):545-548
Individual variability in sucrose consumption is prominent in humans and other species. To investigate the genetic contribution
to this complex behavior, we conducted behavioral, electrophysiological, and genetic studies, using male progeny of two inbred
mouse strains (C57BL/6ByJ [B6] and 129/J [129]) and their F2 hybrids. Two loci on Chromosome (Chr) 4 were responsible for over 50% of the genetic variability in sucrose intake. These
loci apparently modulated intake by altering peripheral neural responses to sucrose. One locus affected the response threshold,
whereas the other affected the response magnitude. These findings suggest that the majority of difference in sucrose intake
between male B6 and 129 mice is due to polymorphisms of two genes that influence receptor or peripheral nervous system activity.
Received: 27 January 1997 / Accepted: 17 March 1997 相似文献
224.
225.
Expression of connexin43 mRNA in adult rodent brain 总被引:3,自引:0,他引:3
E. Simbürger Alexander Stang Marian Kremer R. Dermietzel 《Histochemistry and cell biology》1997,107(2):127-137
226.
The neoglycolipid technology comprises several microproceduresinvolving the generation of lipid-linked oligosaccharide probesfor carbohydrate recognition studies in conjunction with oligosaccharidesequence determination by mass spectrometry. Although applicableto any desired oilgosaccharides, procedures are greatly facilitatedif the ohgosaccharides are nonreduced, as conjugation is byreductive amination of a reducing end aldehyde to a phosphatidylethanolamine.Using bovine submaxillary mucin as a model for release of O-glycansin the reducing state, and based on yields of neoglycolipidsand side-products from "peeling" reactions and degradation,aqueous ethylamine 70% w/v at 22°C for 48 h has been selectedin preference to other conditions, triethylainine, sodium hydroxide,and bydrazine. The integrity of the main acidic and neutraloligosaccharides released under these conditions, di- to octasaccharides,was established by analyses of free oligosaccharides by liquidsecondary ion mass spectrometry (LSIMS) and of the derived neoglycolipidsby TLCLSIMS; the repertoire compared favorably with that ofthe oligosaccharide alditols generated by conventional reductivealkaline borohydride treatment. More forcing conditions of ethylamine70% w/v at 65°C for 6 h were required to release oligosaccharidesfrom porcine gastric mucin; di- to nonasaccharides were obtainedof which about one-third had an intact core GalNAc. Relativeto yields after reductive alkaline hydrolysis, the overall yieldsfor these two glycoproteins were 20% and 4050% for acidicand neutral oligosaccharides, respectively. Among O-glycansreleased from an ovarian cystadenoma glycoprotein using ethylamine,three variants of the sulfated Lea/x sequences were identifiedas ligands for the endothelial adhesion molecule E-selectin,one of which is based on the unusual backbone Gal-3/4GlcNAc-3Gal-3Gal. mucins O-linked oligosaccharides TLC-LSIMS neoglycolipids E-selectin 相似文献
227.
228.
Extensive Sequence Conservation Among Insect,Nematode, and Vertebrate Vitellogenins Reveals Ancient Common Ancestry 总被引:10,自引:0,他引:10
Jeng-Shong Chen Thomas W. Sappington Alexander S. Raikhel 《Journal of molecular evolution》1997,44(4):440-451
The eggs of most oviparous animals are provisioned with a class of protein called vitellogenin (Vg) which is stored as the
major component of yolk. Until recently, deduced amino acid sequences were available only from vertebrate and nematode Vgs,
which proved to be homologous. The sequences of several insect Vgs are now known, but early attempts at pairwise alignments
with vertebrate and nematode Vgs have been problematic, leading to conflicting conclusions about how closely insect Vgs are
related to the others. In this paper we demonstrate that insect Vg sequences can be confidently aligned with one another along
their entire lengths and with multiple vertebrate and nematode Vg sequences along most of their spans. Although divergence
is high, conservation among insect, vertebrate, and nematode Vg sequences is widespread with a preponderance of glycine, proline,
and cysteine residues among strictly conserved amino acids, establishing conclusively that Vgs from the three phyla are homologous.
Areas of least-certain alignment are primarily in and around insect and vertebrate polyserine domains which are not homologous.
Phylogenetic reconstructions of Vgs based on sequence identities indicate that the insect lineage is the most diverged and
that the mammalian serum protein, apolipoprotein B-100, arose from a Vg ancestor after the nematode/vertebrate divergence.
Received: 6 May 1996 / Accepted: 27 September 1996 相似文献
229.
The reproductive biology and phycobiliproteins of four different culture isolates of the freshwater algae Audouinella and‘Chantransia’were investigated.‘Chantransia’sp. (3585/UTEX 2623) and Audouinella macrospora (Wood) Sheath et Burkholder (3394,3395) from California and Minnesota reproduced only by monospores. However, A. macrospora (3603/Necchi 1) reproduced by monosporangia that formed successive generations of the Audouinella phase, and Batrachospermum shoots developed from the basal and erect systems. The major light-harvesting phycobiliprotein in all of these isolates was a phycocyanobilin-containing phycoerythrin not previously detected in red algae or cyanobacteria. As in the commonly found R- and B-phycoerythrins, Audouinella phycoerythrin had a native molecular mass of ~ 240,000 and was made up of α, β, and γ subunits. Audouinella phycoerythrin carried two phycoerythrobilins on the α subunit; one phycourobilin, one phycoerythrobilin, and one phycocyanobilin on the β subunit; and one phycourobilin and two phycoerythrobilins on the γ subunit. With excitation at 495, 563, or 603 nm, the fluorescence emission peak of Audouinella phycoerythrin was at 626 nm, showing that phycocyanobilin was the terminal energy acceptor. 相似文献
230.
A role for natural simian immunodeficiency virus and human immunodeficiency virus type 1 nef alleles in lymphocyte activation. 总被引:7,自引:2,他引:5 下载免费PDF全文
A T-lymphoid cell line termed 221 was derived from a rhesus monkey infected with herpesvirus saimiri. Growth of 221 cells was dependent on the addition of interleukin-2 (IL-2) to the culture medium. In the absence of IL-2, 221 cells arrested in G0-G1 but did not die. Simian immunodeficiency virus (SIV) replicated efficiently in IL-2-stimulated 221 cells whether or not the nef gene was present. In the absence of IL-2, nef-containing SIV replicated 8 to 100 times more efficiently in 221 cells than did the same virus lacking nef. nef-containing virus preferentially stimulated the production of IL-2 from 221 cells. HIV-1 nef and v-ras genes, but not the c-ras gene, were shown to substitute functionally for SIV nef when tested as recombinant viruses in this assay system. These results demonstrate a role for natural nef in causing lymphoid cell activation, and they provide a system for delineating the biochemical mechanisms responsible for this activation. 相似文献