Abstract：
:A newly recognized type of protein-ligand interaction phenomenon has resulted in the preparation of simple, nonionic, and highly specific gel derivatives for selective adsorption chromatography. The essential structure of the immobilized ligand can be represented as agarose-CH2CH2SO2CH2CH2SCH2CH2OH, which was prepared by using mercaptoethanol to derivatize [0.9-1.0 mmol (g of dry gel)-1] divinyl sulfone activated agarose (thiophilic or T-gel). Proteins interacting with this ligand are provisionally termed "thiophilic" to recognize their affinity for the definitive sulfone-thioether constituents. To better understand the experimental variables affecting adsorption efficiency and selectivity, several well-characterized proteins with diverse physicochemical features have been evaluated for thiophilic properties. Thiophilic interaction chromatography was investigated as a function of pH as well as the type and concentration of water-structure-forming salts required to promote adsorption. The model proteins characterized varied distinctly in their individual thiophilic affinities. At acidic pH values, a salt-independent adsorption process was observed. Furthermore, a minimum in the salt-promoted thiophilic adsorption tendency at pH 5-6 was found, with varying magnitude, for each of the model proteins evaluated. Recovery of adsorbed proteins routinely varied from 90% to 100%. There does not appear as yet to be any easily recognized physicochemical property associated with either thiophilic or nonthiophilic behavior. These results suggest that thiophilic interaction chromatography is a process that utilizes a previously unrecognized protein-ligand interaction mechanism. We suggest that salt allows the protein into close proximity with the sulfone-thioether group where short-range forces are effective.(ABSTRACT TRUNCATED AT 250 WORDS)
journal_name
Biochemistryjournal_title
Biochemistryauthors
Hutchens TW,Porath Jdoi
10.1021/bi00396a049subject
Has Abstractpub_date
1987-11-03 00:00:00pages
7199-204issue
22eissn
0006-2960issn
1520-4995journal_volume
26pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract：:Klebsiella aerogenes urease uses a dinuclear nickel active site to catalyze the hydrolysis of urea. Here, we describe the steady-state and pre-steady-state kinetics of urease inhibition by fluoride. Urease is slowly inhibited by fluoride in both the presence and absence of substrate. Steady-state rate studies yield pa...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi992287m
更新日期：2000-05-09 00:00:00
abstract：:The secondary structure of protein kinase C alpha (PKC alpha) has been studied using infrared spectroscopy in the presence of both H(2)O and D(2)O buffers. In the absence of ligands at 20 degrees C, it was shown that beta-sheet is the major component, representing about 44% of the total structure, whereas the alpha-he...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi035128i
更新日期：2004-03-02 00:00:00
abstract：:The steady-state tryptophan fluorescence and time-resolved tryptophan fluorescence of Escherichia coli thioredoxin, calf thymus thioredoxin, and yeast thioredoxin have been studied. In all proteins, the tryptophan residues undergo strong static and dynamic quenching, probably due to charge-transfer interactions with t...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00434a038
更新日期：1989-04-18 00:00:00
abstract：:Glutaminase (GLS1/2) catalyzes the conversion of L-glutamine to L-glutamate and ammonia. The level of a splice variant of GLS1 (GAC) is elevated in certain cancers, and GAC is specifically inhibited by bis-2-(5-phenylacetimido-1,2,4,thiadiazol-2-yl)ethyl sulfide (BPTES). We report here the first full-length crystal st...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi201613d
更新日期：2011-12-20 00:00:00
abstract：:Four different temperature-sensitive M protein mutants (tsM) of vesicular stomatitis virus (VSV) were characterized with regard to the association of the mutated M protein either with nucleocapsids or with membranes in the intact virions. Virions were labeled with the photoreactive hydrophobic probe [125I]iodonaphthyl...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00527a020
更新日期：1981-11-24 00:00:00
abstract：:Esterification of retinol (vitamin A alcohol) with long-chain fatty acids by lecithin-retinol acyltransferase (LRAT) is an important step in both the absorption and storage of vitamin A. Retinol in cells is bound by either cellular retinol binding protein (CRBP), present in most tissues including liver, or cellular re...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00144a014
更新日期：1992-07-28 00:00:00
abstract：:We have employed a combination of gel retardation, protein-DNA cross-linking, and protein-protein cross-linking techniques to further examine the 2,3,7,8-tetrachlorodibenzo-p- dioxin-(TCDD-) dependent changes in the Ah receptor that result in a DNA-binding conformation. Gel retardation analysis of DNA-Sepharose chroma...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00225a026
更新日期：1991-03-19 00:00:00
abstract：:12-Iodo-cis-9-octadecenoic acid (12-IODE) is a time-dependent, irreversible inactivator of soybean lipoxygenase 1. The rate of inactivation is independent of 12-IODE concentration above 20 microM and is half-maximal at about 4 microM. Inactivation by 12-IODE requires lipid hydroperoxide, which must be present even aft...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00424a019
更新日期：1988-11-29 00:00:00
abstract：:Earlier thermodynamic studies of the intermolecular interactions between mature oxytocin and neurophysin, and of the effects of these interactions on neurophysin folding, raised questions about the intramolecular interactions of oxytocin with neurophysin within their common precursor. To address this issue, the disulf...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi9912950
更新日期：1999-10-12 00:00:00
abstract：:Type VI collagen, prepared from guanidine extracts of human amnion, contains very little monomeric material, the major forms being dimers and tetramers. In order to study the orientation of the molecules in these aggregates, they were digested with pepsin followed by bacterial collagenase. Two fragments were isolated,...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00435a020
更新日期：1989-05-02 00:00:00
abstract：:Triton X-100 solubilized monomers of bacteiorhodopsin (bR) show a decrease in the extent of light adaptation; the red shift and the absorbance increase of the visible absorption band are reduced no less than half the values observed in purple membrane (p.m.) with a corresponding reduction in the isomerization of 13-ci...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00555a043
更新日期：1980-07-08 00:00:00
abstract：:Manganese peroxidase (MnP) from Phanerochaete chrysosporium undergoes a pH-dependent conformational change evidenced by changes in the electronic absorption spectrum. This high- to low-spin alkaline transition occurs at approximately 2 pH units lower in an F190I mutant MnP when compared to the wild-type enzyme. Herein...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi000679j
更新日期：2000-08-15 00:00:00
abstract：:Deamidation of proteins is one of the most prevalent post-translational modifications found upon aging, and in age-onset diseases. Specific asparagine and glutamine residues are often selectively deamidated during this process. In the human lens, deamidation has been shown to occur in many crystallins, but it is not c...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/acs.biochem.5b00185
更新日期：2015-08-11 00:00:00
abstract：:To gain insight into the folding of large proteins, we constructed a bovine beta-lactoglobulin (beta-lg) dimeric mutant, A34C/C121A beta-lg. In the mutant, a free thiol group of wild-type beta-lg at Cys121 was removed and two beta-lg molecules were linked by a disulfide bridge through Cys34 created at the dimer's inte...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi8001715
更新日期：2008-06-03 00:00:00
abstract：:Thymidylate synthase from phage T4 (T4TS) is part of a complex of several enzymes required for coordinate DNA synthesis in infected Escherichia coli cells. It has been proposed that similar complexes of enzymes related to DNA synthesis are also functional in eukaryotes [Pardee, A. B. (1989) Science 246, 603-608]. To d...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00255a028
更新日期：1994-12-27 00:00:00
abstract：:Mixtures of 1,2-dipalmitoylglycerol (1,2-DPG), 1,2-dioleylglycerol (1,2-DOG), 1,2-dicapryloylglycerol (1,2-DCG), 1,3-dioleylglycerol (1,3-DOG), and 1,3-dicapryloylglycerol (1,3-DCG) with dipalmitoylphosphatidylcholine (DPPC) have been studied by means of differential scanning calorimetry (DSC) and fluorescence depolar...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00425a022
更新日期：1988-12-13 00:00:00
abstract：:A dark state tertiary structure in the cytoplasmic domain of rhodopsin is presumed to be the key to the restriction of binding of transducin and rhodopsin kinase to rhodopsin. Upon light-activation, this tertiary structure undergoes a conformational change to form a new structure, which is recognized by the above prot...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi010746p
更新日期：2001-10-23 00:00:00
abstract：:Two dioxygenases (ARD and ARD') were cloned from Klebsiella pneumoniae that catalyze different oxidative decomposition reactions of an advanced aci-reductone intermediate, CH(3)SCH(2)CH(2)COCH(OH)=CH(OH) (I), in the methionine salvage pathway. The two enzymes are remarkable in that they have the same polypeptide seque...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi010110y
更新日期：2001-05-29 00:00:00
abstract：:Covalent labeling of the canine renal parathyroid hormone receptor with [125I]bPTH(1-34) reveals several major binding components that display characteristics consistent with a physiologically relevant adenylate cyclase linked receptor. Through the use of the specific glycosidases neuraminidase and endoglycosidase F a...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00398a044
更新日期：1987-12-01 00:00:00
abstract：:The mass-driven assembly of spectrin dimers to form tetramers involves two equal head-to-head alpha-beta associations and requires at least 30 degrees C for interconversion to occur readily. In this paper, the properties of tetramer formation were investigated using two complementary univalent peptides (the alpha I do...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00159a030
更新日期：1992-11-10 00:00:00
abstract：:The heat-stable, protein inhibitor of the cyclic adenosine monophosphate (cAMP) dependent protein kinase [Walsh, D. A., Ashby, C. D., Gonzalez, C., Calkins, D., Fischer, E., & Krebs, E (1971a) J. Biol. Chem. 246, 1977-1985] has been purified to homogeneity from rabbit skeletal muscle by preparative electrophoresis. Em...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00589a011
更新日期：1979-10-30 00:00:00
abstract：:The stable tyrosine radical YD. (tyrosine 160 in the D2 polypeptide) in photosystem II (PSII) exhibits nonexponential electron spin-lattice relaxation transients at low temperature. As previously reported, the tetranuclear Mn complex in PSII significantly enhances the spin-lattice relaxation of YD.. However, in Mn-dep...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00117a033
更新日期：1992-01-21 00:00:00
abstract：:Dihydrouridine is absent from the tRNA of almost all hyperthermophiles and most Archaea but is ubiquitous in the tRNA of Eubacteria and Eukaryotes. In order to investigate whether this could be due to instability, the rate of ring opening of dihydrouridine was measured between 25 and 120 degrees C. The dihydrouridine ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi951577+
更新日期：1996-01-09 00:00:00
abstract：:The crystal structure of a ternary complex of the alcohol dehydrogenase from the archaeon Sulfolobus solfataricus (SsADH) has been determined at 2.3 A. The asymmetric unit contains a dimer with a NADH and a 2-ethoxyethanol molecule bound to each subunit. The comparison with the apo structure of the enzyme reveals that...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi035271b
更新日期：2003-12-16 00:00:00
abstract：:The complete amino acid sequence of human liver cytosolic alanine aminotransferase (GPT) (EC 2.6.1.2) is presented. Two primary sets of overlapping fragments were obtained by cleavage of the pyridylethylated protein at methionyl and lysyl bonds with cyanogen bromide and Achromobacter protease I, respectively. Isolated...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00107a013
更新日期：1991-10-29 00:00:00
abstract：:The structures of beta class carbonic anhydrases (beta-CAs) determined so far fall into two distinct subclasses based on the observed coordination of the catalytic zinc (Zn2+) ion. The subclass of beta-CAs that coordinate Zn2+ tetrahedrally with four protein-derived ligands is represented by the structures of ortholog...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi052272q
更新日期：2006-04-11 00:00:00
abstract：:The temperature dependence of the 31P NMR spectra of yeast phenylalanine tRNA, E. coli tyrosine, glutamate (2), and formylmethionine tRNA is presented. The major difference between the 31P NMR spectra of the different acceptor tRNAs is in the main cluster region between -0.5 and -1.3 ppm. This confirms an earlier assi...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00266a018
更新日期：1982-11-09 00:00:00
abstract：:The Rhodnius prolixus aggregation inhibitor 1 (RPAI-1) is a novel blood-sucking salivary molecule that binds to ADP and attenuates platelet aggregation. In this report, we determine the binding constants and specificity of RPAI-1 for adenine nucleotides and its functional significance. By the Hummel-Dreyer method of e...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi011015s
更新日期：2002-03-19 00:00:00
abstract：:In addition to its superoxide dismutase (SOD) activity, Cu,Zn-superoxide dismutase (CuZnSOD) catalyzes the reductive decomposition of S-nitroso-L-glutathione (GSNO) in the presence of thiols such as L-glutathione (GSH). The GSNO-reductase activity but not the superoxide dismutase (SOD) activity of CuZnSOD is inhibited...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi061077o
更新日期：2006-10-24 00:00:00
abstract：:The 14-3-3 proteins interact with diverse cellular molecules involved in various signal transduction pathways controlling cell proliferation, transformation, and apoptosis. To aid our investigation of the biological function of 14-3-3 proteins, we have set out to identify high-affinity antagonists. By screening phage ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi991353h
更新日期：1999-09-21 00:00:00
