Abstract:
:Human fibroblast interferon binds to a concanavalin A-agarose (Con A-Sepharose) equilibrated with methyl alpha-D-mannopyranoside, or levan; in contrast, it is only partially retarded on a similar column equilibrated with ethylene glycol. Interferon does not bind, however, to a lectin column equilibrated with both methyl alpha-D-mannopyranoside and ethylene glycol. Thus, a hydrophobic interaction between fibroblast interferon and the immobilized lectin seems to account for a large portion of the binding forces involved. Other hydrophobic solutes, such as dioxane, 1, 2-propanediol, and tetraethylammonium chloride, were found equally or more efficient than ethylene glycol in displacing interferon from the lectin column. The elution pattern of interferon from a concanavalin A-agarose (Con A-Sepharose) column, at a constant ehtylene glycol concentration and with an increasing mannoside concentration, reveals the existence of four distinct interferon components. The selective adsorption to and elution from a concanavalin A-agarose (Con A-Sepharose) column resulted in about a 3000-fold purification of human fibroblast interferon and complete recovery of activity. The specific activity of the partially purified interferon preparation is about 5 X 10(7) units per mg of protein. The chromatographic behavior of human leukocyte interferon is remarkable in that it does not bind to concanavalin A-agarose at all indicating the absence of carbohydrate moieties recognizable by the lectin, or if present, their masked status. When concanavalin A was coupled to an agarose matrix (cyanogen bromide activated) at pH 8.0 and 6.0 human fibroblast interferon bound to both lectin-agarose adsorbents and could be recovered with methyl alpha-D-mannopyranoside. Concanavalin A, immobilized directly on agarose matrix at pH 8.0 and 6.0, thus displays only carbohydrate recognition toward interferon. By contrast, unless a hydrophobic solute was included in the solvent containing methyl mannoside, human fibroblast interferon could not be recovered from concanavalin A-agarose coupled at pH 9.0. When concanavalin A was immobilized via molecular arms, in tetrameric as well as dimeric forms, the binding of interferon again occurred exclusively through carbohydrate recognition. Thus, the hydrophobic interaction can be eliminated by appropriate immobilization of the lectin, and then adsorbed glycoproteins, as exemplified here by interferon, can be recovered readily with methyl mannoside alone.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Davey MW,Sulkowski E,Carter WAdoi
10.1021/bi00648a039subject
Has Abstractpub_date
1976-02-10 00:00:00pages
704-13issue
3eissn
0006-2960issn
1520-4995journal_volume
15pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract::We introduced a stop codon in place of the ATT codon encoding Ile283 (numbered from the Met initiation codon) in the petA gene from Chlamydomonas reinhardtii. The resulting protein was expected to be truncated on its carboxy-terminus end, lacking the last 35 amino acids. This region of the polypeptide sequence encompa...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00022a021
更新日期:1995-06-06 00:00:00
abstract::We have used DNase I footprinting to examine the binding of five different 17-mer oligonucleotides to a 53-base oligopurine tract containing four pyrimidine interruptions. Although all the expected triplexes formed with high affinity (K(d) approximately 10-50 nM), one oligonucleotide produced a footprint at a second s...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi992773+
更新日期:2000-06-06 00:00:00
abstract::We have measured the concentration of mRNAs coding for immunoglobulins, k and lambda type light chains and gamma 1 type heavy chain, in mouse spleen cells activated by bacterial lipopolysaccharide or sheep red blood cells. These mRNAs were quantitated by hybridization to radioactive DNA complementary to highly purifie...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00645a018
更新日期:1977-12-27 00:00:00
abstract::The mobility of vesicular stomatitis virus (VSV) G protein on the surface of infected BHK cells was studied by using the technique of fluorescence photobleaching recovery. The fraction of surface G protein that was mobile in that time scale of the measurement (minutes) was at least 75%, a relatively high value among c...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00508a047
更新日期:1981-03-03 00:00:00
abstract::Intracellular retinoid-binding proteins are small, tightly folded, compact proteins, which appear to be involved in the delivery of retinoids to microsomal metabolic enzymes, among other potential roles. Recently, it has been demonstrated that two of these binding proteins, cellular retinol-binding protein (CRBP) and ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00176a017
更新日期:1994-03-15 00:00:00
abstract::The synthesis of sphingomyelin was studied in baby hamster kidney cells and in subcellular fractions derived from rat liver. During pulse-chase experiments with [3H]choline in tissue culture cells, the specific radioactivity of sphingomyelin continued to increase after the specific activities of phosphocholine and cyt...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00540a027
更新日期:1982-05-25 00:00:00
abstract::Escherichia coli helicase II (UvrD) protein can initiate unwinding of duplex DNA at blunt ends or nicks, although these reactions require excess protein. We have undertaken kinetic studies of these reactions in order to probe the mechanism of initiation of unwinding. DNA unwinding was monitored directly by using agaro...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00066a039
更新日期:1993-04-20 00:00:00
abstract::We have examined the competitive binding of oxygen and carbon monoxide to the multisubunit hemocyanin of the tarantula Eurypelma californicum. Employment of high-precision thin-layer methods has enabled detailed characterization of the pure oxygen and pure carbon monoxide binding curves, as well as binding curves perf...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00418a036
更新日期:1988-09-06 00:00:00
abstract::Aquifex aeolicus 3-deoxy-d-manno-octulosonate 8-phosphate synthase (KDO8PS) is active with a variety of different divalent metal ions bound in the active site. The Cd(2+), Zn(2+), and Cu(2+) substituted enzymes display similar values of k(cat) and similar dependence of K(m)(PEP) and K(m)(A5P) on both substrate and pro...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi801955h
更新日期:2009-04-28 00:00:00
abstract::Nicotinic acetylcholine receptors (nAChRs) were studied in detail in the past regarding their interaction with therapeutic and drug addiction related compounds. Using fast kinetic whole-cell recording, we have now studied effects of tacrine, an agent used clinically to treat Alzheimer's disease, on currents elicited b...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi101789y
更新日期:2011-03-22 00:00:00
abstract::Ornithine decarboxylase (ODC) is a pyridoxal 5'-phosphate (PLP) dependent homodimeric enzyme. It is a recognized drug target against African sleeping sickness, caused by Trypanosoma brucei. One of the currently used drugs, alpha-difluoromethylornithine (DFMO), is a suicide inhibitor of ODC. The structure of the T. bru...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9915115
更新日期:1999-11-16 00:00:00
abstract::The respiratory chain in the cytochrome d deficient mutant Escherichia coli GR19N is a relatively simple, linear system consisting of primary dehydrogenases, ubiquinone 8, cytochrome b-556, and cytochrome o oxidase. By use of right-side-out and inside-out membrane vesicles from this strain, various oxidase activities ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00357a004
更新日期:1986-05-06 00:00:00
abstract::Purple membranes of Halobacterium halobium were studied by differential scanning calorimetry. No transition was detected at temperatures below 70 degrees C. A small endothermic transition was seen at about 80 degrees C and a larger one at 100 degrees C. The larger transition is the irreversible denaturation of bacteri...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00598a026
更新日期:1978-03-07 00:00:00
abstract::Nitrogenase reduces N2 to NH3, but the mechanistic details are unclear. Diazene (N2H2), a proposed 2e-/2H+ intermediate on the reduction pathway, is labile under typical enzyme assay conditions, and no firm evidence is available on whether or not it can be reduced by or inhibit nitrogenase. In this paper, we compare t...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi950964g
更新日期:1996-04-09 00:00:00
abstract::The existence of two conformational states of concanavalin A (Con A) with different metal ion binding properties has been recently demonstrated (Brown, R. D., Brewer, C. F., & Koenig, S. H. (1977) Biochemistry 16, 3883). Introduction of Mn2+ to the S1 site and Ca2+ to the S2 site of apo-Con A was shown to induce a con...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00613a022
更新日期:1978-10-03 00:00:00
abstract::The Caenorhabditis elegans Werner syndrome protein, WRN-1, a member of the RecQ helicase family, has a 3'-5' DNA helicase activity. Worms with defective wrn-1 exhibit premature aging phenotypes and an increased level of genome instability. In response to DNA damage, WRN-1 participates in the initial stages of checkpoi...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi200791p
更新日期:2012-02-21 00:00:00
abstract::Members of the ATP-binding cassette (ABC) transporters share the same basic architecture, with a four-core domain made of two transmembrane plus two nucleotide-binding domains. However, a supramolecular organization has been detected in some ABC transporters, which might be relevant to physiological regulation of subs...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0482809
更新日期:2005-03-22 00:00:00
abstract::The technique of hydrogen-deuterium exchange coupled to mass spectrometry (HDX-MS) has been applied to a mesophilic (E. coli) dihydrofolate reductase under conditions that allow direct comparison to a thermophilic (B. stearothermophilus) ortholog, Ec-DHFR and Bs-DHFR, respectively. The analysis of hydrogen-deuterium e...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi200640s
更新日期:2011-09-27 00:00:00
abstract::Phytate is an antinutritional factor that influences the bioavailability of essential minerals by forming complexes with them and converting them into insoluble salts. To further our understanding of the chemistry of phytate's binding interactions with biologically important metal cations, we determined the stoichiome...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi1010249
更新日期:2010-11-30 00:00:00
abstract::Recently, a new clathrin assembly protein (AP180) has been purified from coated vesicles of bovine brain (Ahle & Ungewickell, 1986). This protein has been shown to promote polymerization of clathrin into a homogeneous population of baskets under conditions where pure clathrin does not polymerize by itself. We have pur...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00416a040
更新日期:1988-08-09 00:00:00
abstract::S-Adenosylmethionine decarboxylase is a pyruvate-dependent enzyme. The enzyme forms a Schiff base with substrate, S-adenosylmethionine, through the pyruvoyl moiety. This facilitates the release of CO2 from the substrate, which must then be protonated on the alpha carbon in order to permit hydrolysis of the Schiff base...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9825201
更新日期:1999-02-23 00:00:00
abstract::Microfibrils having a diameter of 10-12 nm, found either in association with elastin or independently, are an important component of the extracellular matrix of many tissues. Because isolation of native proteins composing the microfibrils has proven difficult, information on structure/function relationships is limited...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00168a026
更新日期:1994-01-18 00:00:00
abstract::These investigations characterize the covalent binding of reactive products of prostaglandin H-synthases (PGHSs) to the enzyme and to other molecules. The intermediate product of oxygenation of arachidonic acid by the PGHSs, prostaglandin (PG) H2, undergoes rearrangement to the highly reactive gamma-keto aldehydes, le...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi002629k
更新日期:2001-06-12 00:00:00
abstract::Polylysine promoted extensive membrane mixing of liposomes only if the buffer pH was below the pKa of the lysyl residues. This observation suggested that fusion could be regulated in a physiological pH range if the homopolymer of L-histidine was substituted as fusogen. Microgram quantities of polyhistidine were added ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00322a001
更新日期:1985-01-01 00:00:00
abstract::The equilibrium binding of influenza virus hemagglutinin to derivatives of its cell-surface ligand, sialic acid, was measured by nuclear magnetic resonance (NMR) spectroscopy. Binding was quantified by observing perturbations of sialic acid resonances in the presence of protein. The major perturbation observed was a c...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00447a018
更新日期:1989-10-17 00:00:00
abstract::Autophagy, an essential eukaryotic homeostasis pathway, allows the sequestration of unwanted, damaged, or harmful cytoplasmic components in vesicles called autophagosomes, permitting subsequent lysosomal degradation and nutrient recycling. Autophagosome nucleation is mediated by class III phosphatidylinositol-3-kinase...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.6b00246
更新日期:2016-08-02 00:00:00
abstract::The regiospecificity and stereospecificity of proton transfer in the yeast inorganic pyrophosphatase (PPase) catalyzed hydrolysis of P1,P2-bidentate Mg(H2O)4(PPi)2- were probed with exchange-inert metal complexes of imidodiphosphate (PNP) and thiopyrophosphate (PPS). PPase was unable to catalyze the hydrolysis of Mg(H...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00364a035
更新日期:1986-08-12 00:00:00
abstract::RP2 is a ubiquitously expressed protein encoded by a gene associated with X-linked retinitis pigmentosa (XLRP), a retinal degenerative disease that causes severe vision loss. Previous in vitro studies have shown that RP2 binds to ADP ribosylation factor-like 3 (Arl3) and activates its intrinsic GTPase activity, but th...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi1005249
更新日期:2010-09-07 00:00:00
abstract::Native calf thymus DNA was reacted with N-acetoxy-N-2-acetylaminofluorene (N-AcO-AAF) and its 7-fluoro and 7-iodo derivatives. Different ways of purification of the fluorene modified DNA samples were checked in order ot obtain a nucleic acid free from all noncovalently bound fluorene residues. The decrease in melting ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00660a027
更新日期:1976-07-27 00:00:00
abstract::Identical first-order rate constants for phosphorylation of the calcium ATPase of sarcoplasmic reticulum by bound inorganic phosphate (Pi) of 25 +/- 2 s-1 with empty vesicles and 25 +/- 1 s-1 with vesicles that were passively loaded with 40 mM Ca2+ were obtained by treating the reaction as an approach to equilibrium (...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00009a039
更新日期:1995-03-07 00:00:00