Abstract：
:A bifunctional molecule was genetically engineered which contained an amino-terminal effector domain that bound immunoglobulin Fc (fragment B of staphylococcal protein A) and a carboxyl-terminal domain that bound digoxin [a single-chain Fv (sFv)]. Effector and sFv binding properties were virtually identical with those of the parent molecules, despite the proximity of the FB to the sFv combining site. This finding is unprecedented since in all molecules of the natural immunoglobulin superfamily, the antigen binding domain is amino terminal to the effector domain. The FB-sFv sequence was encoded in a single synthetic gene and expressed as a 33,106 molecular weight protein in Escherichia coli. After purification, renaturation, and affinity isolation, yield of active fusion protein were 110 mg/L of fermented cells (18.5-g cell paste). Bifunctionality was confirmed by the ability of FB-sFv to cross-link IgG to digoxin-bovine serum albumin, as measured by plate assays and by Ouchterlony analysis. Analysis of the expressed fusion protein suggests that the sFv holds promise for the development of multifunctional, targetable single-chain proteins.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Tai MS,Mudgett-Hunter M,Levinson D,Wu GM,Haber E,Oppermann H,Huston JSdoi
10.1021/bi00487a005subject
Has Abstractpub_date
1990-09-04 00:00:00pages
8024-30issue
35eissn
0006-2960issn
1520-4995journal_volume
29pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract：:An inhibitor of the insulin receptor tyrosine kinase (IRTK), (hydroxy-2-naphthalenyl-methyl) phosphonic acid, was designed and synthesized and was shown to be an inhibitor of the biological effects of insulin in vitro. With a wheat germ purified human placental insulin receptor preparation, this compound inhibited the...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00439a053
更新日期：1989-06-27 00:00:00
abstract：:Photosystem II catalyzes the light-driven oxidation of water and reduction of plastoquinone in oxygenic photosynthesis. The manganese stabilizing protein (MSP) of photosystem II is an extrinsic subunit that plays an important role in catalytic activity. This subunit can be extracted and re-bound to the photosystem II ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi9724467
更新日期：1998-04-21 00:00:00
abstract：:The C-terminal domain from the muscle protein troponin C (TnC) comprises two helix-loop-helix calcium-binding sites (residues 90-162). The assembly of these two sites is governed by calcium binding enabling a synthetic C-terminal domain to be preferentially and stoichiometrically assembled from two synthetic peptides ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi9528006
更新日期：1996-06-11 00:00:00
abstract：:Escherichia coli were grown on 14.3% uniformly 13C-labeled glucose as the sole carbon source and challenged anaerobically with 90% 13C-labeled formaldehyde. The major multiply labeled metabolites were identified by 13C NMR spectroscopy to be glycerol and 1,2-propanediol, and a minor metabolite was shown to be 1,3-prop...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00336a050
更新日期：1985-07-16 00:00:00
abstract：:Although the short-chain dehydrogenase/reductase (SDR) superfamily contains a very large number of members defined in annotated databases and by biochemical and structural studies, very few SDR enzymes have been identified that have a homologous partner catalyzing the same reaction but with an opposite stereospecifici...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi0118005
更新日期：2002-02-26 00:00:00
abstract：:Mammalian phospholipase D (PLD) has been implicated in the cellular signal transduction pathways leading to diverse physiological events and known to be regulated by many cellular factors. To identify the proteins that interact with PLD, we performed a protein overlay assay with fractions obtained from the sequential ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi015700a
更新日期：2002-03-12 00:00:00
abstract：:Frequency-resolved fluorescence measurements have been performed to quantitate the lateral stress of the lipid layer containing nonbilayer phase preferring dioleoylphosphatidylethanolamine (DOPE). On the basis of a new rotational diffusion model, the wobbling diffusion constant (Dw), the curvature-related hopping diff...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00130a005
更新日期：1992-04-21 00:00:00
abstract：:Human S100A7 (psoriasin) is a metal-chelating protein expressed by epithelial cells. It is a 22-kDa homodimer with two EF-hand domains per subunit and two transition-metal-binding His3Asp sites at the dimer interface. Each subunit contains two cysteine residues that can exist as free thiols (S100A7red) or as an intram...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/acs.biochem.7b00781
更新日期：2017-10-31 00:00:00
abstract：:Isothermal titration calorimetry (ITC) is a powerful technique for investigating self-association processes of protein complexes and was expected to reveal quantitative data on peroxiredoxin oligomerization by directly measuring the thermodynamic parameters of dimer-dimer interaction. Recombinant classical 2-cysteine ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi8002956
更新日期：2008-07-08 00:00:00
abstract：:The effects of poly(ethylene glycol) (PEG) on sonicated unilamellar vesicles made of phosphatidylcholine have been examined. Stopped-flow and equilibrium data are presented for vesicle aggregation, vesicle leakage, lipid mixing, and aqueous contents mixing. Vesicle aggregation is detected as a monoexponential increase...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00065a024
更新日期：1993-04-13 00:00:00
abstract：:The dietary flavonoid quercetin has a broad range of biological activities, including potent antitumor activity against several types of tumors. Recently, it has been shown that quercetin inhibits cancer cells proliferation by depleting cellular microtubules and perturbing cellular microtubule functions. However, the ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi025952r
更新日期：2002-10-29 00:00:00
abstract：:We have previously designed and synthesized small-molecule inhibitors that reduce Vibrio cholerae virulence in vitro by targeting the transcription factor ToxT. Here we report the synthesis and biological activity of derivatives of our previous bicyclic, fatty acid-like inhibitors. All of the synthesized derivatives s...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/acs.biochem.8b00667
更新日期：2018-09-25 00:00:00
abstract：:The active form of the animal fatty acid synthase (FAS) is a dimer of identical multifunctional polypeptides, each containing seven discrete functional domains, that cooperate to form two centers for palmitate synthesis. To assess the importance of domain cooperation across the subunit interface in the reaction mechan...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi971886v
更新日期：1998-02-24 00:00:00
abstract：:The Raman spectra of the nonbridging V--O bonds in the myosin S1.MgADP.Vi complex, often believed to be a transition-state analogue for the phosphotransfer reaction catalyzed by myosin, and in a vanadate solution model compound have been obtained using Raman difference spectroscopic techniques. A symmetric/asymmetric ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi980556n
更新日期：1998-08-04 00:00:00
abstract：:The reactivity of a series of para-substituted phenolic compounds in the peroxidation catalyzed by chloroperoxidase was investigated, and the results were interpreted on the basis of the binding characteristics of the substrates to the active site of the enzyme. Marked selectivity effects are observed. These operate t...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00187a001
更新日期：1994-05-31 00:00:00
abstract：:The SH3 and SH2 domains of hematopoietic cell kinase (Hck) play important roles in substrate targeting. To identify new components of Hck signaling pathways, we identified proteins that bind to the SH3 domain of Hck (Scott et al. (2002) J. Biol. Chem. 277, 28238). One such protein was ELMO1, the mammalian orthologue o...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi0500832
更新日期：2005-06-21 00:00:00
abstract：:Retinoid X receptors (RXRs) are nuclear receptors that can activate transcription as homodimers or as obligate heterodimeric partners of other nuclear receptors. While the crystal structures of the RXR ligand-binding domains (LBD) have been previously determined, the dynamics of activation is less well characterized a...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi051474j
更新日期：2006-02-14 00:00:00
abstract：:Proline-71, an evolutionally conserved residue that separates two short alpha-helical regions, is replaced by valine, threonine, or isoleucine in at least partially functional forms of iso-1-cytochrome c from Saccharomyces cerevisiae [Ernst, J. F., Hampsey, D. M., Stewart, J. W., Rackovsky, S., Goldstein, D., & Sherma...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00370a033
更新日期：1986-11-04 00:00:00
abstract：:The beta- and delta-subunits of the nicotinic acetylcholine receptor from Torpedo californica were covalently photolabeled at the synaptic surface with the ATP photoaffinity analogue [alpha-32P]-8-azido-ATP. The specificity of labeling for nucleotide binding sites was demonstrated by the saturation of labeling with in...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00080a002
更新日期：1993-07-27 00:00:00
abstract：:Subfragment 1 (S1) prepared from rabbit skeletal muscle myosin was digested with trypsin to cleave the 95K heavy chain into three pieces, i.e., the 23K, 50K, and 20K fragments. The trypsin-treated S1 was then cross-linked with p-nitrophenyl iodoacetate. The cross-linker bridged one of the reactive thiols (SH2) in the ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00408a045
更新日期：1988-04-19 00:00:00
abstract：:DB2277, a heterocyclic diamidine, is a successful design for mixed base pair (bp) DNA sequence recognition. The compound has a central aza-benzimidazole group that forms two H-bonds with a GC bp that has flanking AT bps. The nuclear magnetic resonance structure of the DB2277-DNA complex with an AAGATA recognition site...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/acs.biochem.8b00647
更新日期：2018-08-21 00:00:00
abstract：:One-carbon units are essential to a variety of anabolic processes which yield necessary cellular components including purines, pyrimidines, amino acids, and lipids. Serine hydroxymethyltransferase (SHMT) is the major provider of one-carbon units in the cell. The other product of this reaction is glycine. Both of these...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi971610n
更新日期：1997-12-02 00:00:00
abstract：:We report here that polyamide nucleic acid (PNA) as well as a polyamide nucleic acid-DNA chimera complementary to the primer binding site of the HIV-1 genome can completely block priming by tRNA3Lys and consequently the in vitro initiation of reverse transcription by HIV-1 RT. Conventional heating and cooling is not r...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi972197m
更新日期：1998-01-20 00:00:00
abstract：:Apamin is being studied as a model for the folding mechanism of proteins whose structures are stabilized by disulfide bonds. Apamin consists of 18 amino acid residues and forms a stable structure consisting of a C-terminal alpha-helix and two reverse turns. This structure is stabilized by two disulfide bonds connectin...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00133a009
更新日期：1992-05-12 00:00:00
abstract：:Kinesin-5 proteins are essential for formation of a bipolar mitotic spindle in most and, perhaps all, eukaryotic cells. Several Kinesin-5 proteins, notably the human version, HsEg5, are targets of a constantly expanding group of small-molecule inhibitors, which hold promise both as tools for probing mechanochemical tr...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi801291q
更新日期：2009-03-03 00:00:00
abstract：:Human liver aldehyde dehydrogenase isozymes E1 and E2 (EC 1.2.1.3) are both completely and irreversibly inactivated by bromoacetophenone (2-bromo-1-phenylethanone). Steady-state kinetics with both acetophenone and chloroacetophenone indicated interaction with the same enzyme form as the aldehyde substrate. Saturation ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00366a030
更新日期：1986-09-09 00:00:00
abstract：:Solutions of porcine lactate dehydrogenase of micromolar concentration kept at 4 degrees C for several days lose the greater part of their enzymic activity but recover it when returned to room temperature. The rate of spoiling decreases and the rate of recovery increases with the concentration of the solutions. The de...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00360a024
更新日期：1986-06-17 00:00:00
abstract：:Deposition of misfolded alpha-synuclein is a critical factor in several neurodegenerative disorders. Filamentous alpha-synuclein is the major component of Lewy bodies and Lewy neurites, the intracellular inclusions in the dopaminergic neurons of the substantia nigra, which are considered the pathological hallmark of P...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi034938r
更新日期：2004-03-23 00:00:00
abstract：:Here, we biochemically characterize Dictyostelium myosin II mutants that were previously phenotypically selected following random mutagenesis and shown to lie in the actin binding domain [Patterson, B., & Spudich, J. A. (1996) Genetics 143, 801-810]. We show that the conditional loss of myosin-dependent activity in vi...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi963141f
更新日期：1997-07-15 00:00:00
abstract：:Living organisms have evolved a vast array of catalytic functions that make them ideally suited for the production of medicinally and industrially relevant small-molecule targets. Indeed, native metabolic pathways in microbial hosts have long been exploited and optimized for the scalable production of both fine and co...
journal_title：Biochemistry
pub_type： 杂志文章,评审
doi：10.1021/bi200416g
更新日期：2011-06-21 00:00:00
