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 purified this protein from coated vesicles by a simpler method than has been reported. The method involves a gel filtration step on a Sephacryl S-300 column, in 0.5 M Tris-HCl, pH 8.0, and a hydroxylapatite column eluted with 10 mM sodium phosphate/0.5 M Tris-HCl, pH 7.0. By running SDS gels over an extended period of time (5-15% gradient gel, 10 mA for the first 12 h followed by 20 mA for the next 3-4 h) after the marker dye entered the electrode buffer, we have been able to separate AP180 from clathrin heavy chain on the gels. This enabled us to determine its stoichiometry to clathrin heavy chains in isolated coated vesicles and assembled baskets, and was helpful in the purification procedure. The apparent molecular weight of the pure protein on SDS gels was about 180,000, yet gel filtration yielded values of about 120,000. Thus, we undertook the molecular weight determination by another independent method, sedimentation equilibrium analysis, and found a molecular weight of 115,000 and a sedimentation coefficient of 3.50 +/- 0.05 S. Circular dichroism data revealed that it has 30% helical structure, 14% beta-structure, 27% beta-sheet, and the rest random peptides.(ABSTRACT TRUNCATED AT 250 WORDS)
journal_name
Biochemistryjournal_title
Biochemistryauthors
Prasad K,Lippoldt REdoi
10.1021/bi00416a040subject
Has Abstractpub_date
1988-08-09 00:00:00pages
6098-104issue
16eissn
0006-2960issn
1520-4995journal_volume
27pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract::The reaction of mushroom (Agaricus bisporus) tyrosinase with dioxygen in the presence of several o-diphenolic substrates has been studied by steady-state and transient-phase kinetics in order to elucidate the rate-limiting step and to provide new insights into the mechanism of oxidation of these substrates. A kinetic ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi000539+
更新日期:2000-08-29 00:00:00
abstract::Inexpensive, straightforward, and rapid medical diagnostics are becoming increasingly important for disease identification in time- and resource-limited settings. Previous attempts to link oligonucleotide-based aptamers and hammerhead ribozymes to form ligand-induced ribozymes have been successful in identifying a var...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.8b00523
更新日期:2018-08-28 00:00:00
abstract::RNA pur*pur-pyr and pyr*pur-pyr (pur = purine, pyr = pyrimidine) triple helices consisting of a Watson-Crick base-paired 28mer hairpin duplex and a Hoogsteen base-paired purine or pyrimidine 12mer are targeted with photoactivated cleavage by the metal complex Rh(phen)2phi3+ (phen = phenanthroline, phi = 9, 10-phenanth...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi980509v
更新日期:1998-06-23 00:00:00
abstract::The initial step in the l-kynurenine pathway is oxidation of l-tryptophan to N-formylkynurenine and is catalyzed by one of two heme enzymes, tryptophan 2,3-dioxygenase (TDO) or indoleamine 2,3-dioxygenase (IDO). Here, we address the role of the conserved active site Ser167 residue in human IDO (S167A and S167H variant...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi702405a
更新日期:2008-04-22 00:00:00
abstract::The rate of deuterium exchange of a purine 8CH group in DNA is highly sensitive to both macromolecular secondary structure and intermolecular interactions which restrict solvent access to the major groove [Lamba, O.P., Becka, R., & Thomas, G.J., Jr. (1990) Biopolymers 29, 1465-1477]. We have exploited the sensitivity ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00127a012
更新日期:1992-03-31 00:00:00
abstract::Calculations of the electrostatic potentials were made around yeast elongator phenylalanine, aspartate tRNAs, and yeast initiator methionine tRNA in aqueous solution at physiological ionic strength. The calculations were carried out with a finite difference algorithm for solving the nonlinear Poisson-Boltzmann equatio...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00454a006
更新日期:1990-01-16 00:00:00
abstract::Sequence effects on the stability of purine-motif (also called (G, A)-motif) triple helix have been investigated through two symmetry-related systems: one of them had a 5'(GGA)43' core sequence of triplex-forming oligonucleotides (TFOs), whereas the other one had a reversed 5'(AGG)43' core sequence. These (G,A)-contai...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9805588
更新日期:1998-11-24 00:00:00
abstract::The involvement of Cl(-) and several other monovalent anions in photosynthetic oxygen evolution was studied using photosystem II membranes depleted of Cl(-) by dialysis. The results of these studies differ significantly from results obtained using other depletion methods. Binding studies with glycerol as a cryoprotect...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi026175y
更新日期:2003-02-25 00:00:00
abstract::New classes of small proteins have recently been found that refold rapidly with two-state kinetics from a substantially unfolded conformation ("U") and without the accumulation of a folding intermediate. Barnase, on the other hand, is representative of a class of proteins that display multistate kinetics and refold fr...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi950967t
更新日期:1996-02-27 00:00:00
abstract::Treatment of the beta 2 adrenergic receptor with the reducing agent dithiothreitol (DTT) is known to abolish ligand binding to the receptor. Interestingly, the loss of binding can be prevented by preoccupation of the receptor with ligand. It is unclear, however, whether the ligand blocks access of DTT to the receptor,...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi961619+
更新日期:1996-11-19 00:00:00
abstract::Myeloperoxidase plays a fundamental role in oxidant production by neutrophils. The enzyme uses hydrogen peroxide to oxidize chloride (Cl-), bromide (Br-), iodide (I-), and the pseudohalide thiocyanate (SCN-) to their respective hypohalous acids. This study for the first time presents transient kinetic measurements of ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9818772
更新日期:1998-12-22 00:00:00
abstract::In order to gain insight into the structure of human parathyroid hormone (hPTH), four fragments [hPTH(1-34), hPTH(18-48), hPTH(28-48), and hPTH(53-84)], which cover all regions of the intact hormone, have been investigated by CD and NMR spectroscopy in combination with distance geometry, and restrained molecular dynam...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00173a010
更新日期:1994-02-22 00:00:00
abstract::The arrangement of the mRNA on the Escherichia coli ribosome with respect to ribosomal RNA sites has been investigated by photochemical cross-linking experiments. mRNA analogues 51-54 nucleotides in length contained a Shine-Dalgarno sequence, a single codon for tRNA(Gly), and 4-thiouridine (s4U) in the 5' third of the...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00176a040
更新日期:1994-03-15 00:00:00
abstract::The trifunctional dinuclear platinum compounds 1,2/c,c [[cis-PtCl(NH(3))(2)]mu-H(2)N(CH(2))(6)NH(2)[cis-PtCl(2)(NH(3))]](+) and 1,2/t,c [[trans-PtCl(NH(3))(2)]mu-H(2)N(CH(2))(6)NH(2)[cis-PtCl(2)(NH(3))]](+) contain a monofunctional platinum coordination sphere linked to a cis-[PtCl(2)(amine)(2)] moiety. The compounds ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi030243e
更新日期:2004-06-22 00:00:00
abstract::The SfiI restriction enzyme binds to DNA as a tetramer holding two usually distant DNA recognition sites together before cleavage of the four DNA strands. To elucidate structural properties of the SfiI-DNA complex, atomic force microscopy (AFM) imaging of the complexes under noncleaving conditions (Ca2+ instead of Mg2...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi051767c
更新日期:2006-01-10 00:00:00
abstract::DNA sequences with repetitive G-rich structural motifs, which form special structures called G-quadruplexes, widely exist in the human genome. Here we report the general peroxidase activity of G-quadruplex-hemin complexes and discuss the connection between peroxidase activity and G-quadruplex structures. The high pero...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9006786
更新日期:2009-08-25 00:00:00
abstract::Cyanobacteriochromes (CBCRs) are cyanobacterial photoreceptors distantly related to phytochromes. Like phytochromes, CBCRs photointerconvert between two photostates that accompany photoisomerization of their bilin chromophores. While phytochromes typically exhibit red/far-red photocycles, CBCR photocycles are much mor...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi4015538
更新日期:2014-02-18 00:00:00
abstract::Depletion of functional Ca2+ from photosystem (PS) II membranes impairs O2 evolution. Redox properties of the Mn cluster as probed by thermoluminescence were modified differently in Ca(2+)-depleted PSII depending on the procedure for Ca2+ extraction. Ca2+ depletion by low-pH treatment gave rise to an abnormally modifi...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00148a028
更新日期:1992-08-25 00:00:00
abstract::Phenylcarbene and adamantylidene have been generated photochemically from the corresponding diazirines within lipid bilayers. Reasonable yields of labeled fatty acid side chains have been observed. The products have been characterized by gas chromatography-mass spectrometry and derive both from the insertion of the c...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00605a026
更新日期:1978-06-13 00:00:00
abstract::Although the terms "excessive reactive oxygen species (ROS)" and "oxidative stress" are widely used, the implications of oxidative stress are often misunderstood. ROS are not a single species but a variety of compounds, each with unique biochemical properties and abilities to react with biomolecules. ROS cause activat...
journal_title:Biochemistry
pub_type: 杂志文章,评审
doi:10.1021/acs.biochem.6b00703
更新日期:2016-09-27 00:00:00
abstract::We have investigated the topology of the band 3 protein of the human erythrocyte membrane by a combination of chemical labeling and proteolytic cleavage. The N-terminal third of the membrane-bound domain of band 3 is a 17 000-dalton chymotryptic fragment that is known to traverse the membrane an odd number of times. A...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00321a024
更新日期:1984-12-18 00:00:00
abstract::Pyrophosphate-dependent 6-phosphofructo-1-kinase (PPi-PFK) from Propionibacterium freudenreichii was inactivated by low concentrations of the lysine-specific reagent pyridoxal phosphate (PLP) after sodium borohydride reduction. The substrates fructose 6-phosphate and fructose 1,6-bisphosphate protected against inactiv...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00135a011
更新日期:1992-05-26 00:00:00
abstract::The equilibrium binding of hemoglobin to isolated band 3 protein exhibited positive cooperativity [Hill coefficient = 1.65 +/- 0.1; total number of binding sites at pH 6.6 in 5 mM sodium phosphate buffer = 32 500 +/- 940 pmol/mg; Ka = (3.0 +/- 0.5) X 10(5) M-1]. The binding was reversible and ionic in nature as the bo...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00359a054
更新日期:1986-06-03 00:00:00
abstract::Formiminotransferase-cyclodeaminase denatured in 6 M guanidine hydrochloride (Gdn.HCl) refolds and reassembles to the native octameric structure upon dilution into buffer. Both enzymic activities are recovered to greater than 90%, and the renatured enzyme "channels" the formiminotetrahydropteroylpentaglutamate interme...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00409a042
更新日期:1988-05-03 00:00:00
abstract::Calcium is required for the substrate binding and for the chemical step of the interfacial catalytic turnover cycle of pancreatic phospholipase A2 (PLA2), but not for the binding of the enzyme to the interface. The role of calcium and other divalent cations (C) is analyzed for the effect on the substrate binding and k...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9728607
更新日期:1998-09-08 00:00:00
abstract::The subunit structure of Hsp80, the most abundant heat-shock protein of Neurospora crassa, was examined by chemical cross-linking of the purified protein in vitro. Resolution of glutaraldehyde-treated Hsp80 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis SDS-PAGE suggests that the native state of this pro...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi963030g
更新日期:1997-08-19 00:00:00
abstract::Putative physiological partners of the [2Fe-2S] ferredoxin from Clostridium pasteurianum have been searched by running crude soluble extracts of this bacterium through an affinity column to which the [2Fe-2S] ferredoxin had been covalently bound. Subsequent washing of the column with buffers of increasing ionic streng...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi970528p
更新日期:1997-09-30 00:00:00
abstract::Resonance energy-transfer approaches have been used to directly monitor the interactions of the GTP gamma S-bound alpha subunit of transducin (alpha T GTP gamma S) with the retinal cyclic GMP phosphodiesterase (PDE). The PDE was labeled with 5-(iodoacetamido) fluorescein (IAF-PDE) and served as the fluorescence donor ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00243a011
更新日期:1991-07-23 00:00:00
abstract::Multivalent binding is an efficient means to enhance the affinity and specificity of chemical probes targeting multidomain proteins in order to study their function and role in disease. While the theory of multivalent binding is straightforward, physical and structural characterization of bivalent binding encounters m...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.8b00150
更新日期:2018-04-10 00:00:00
abstract::The reactions of fully reduced and mixed-valence bovine heart cytochrome c oxidase with dioxygen have been reinvestigated in the absence and presence of metal ions (Zn(2+), Ni(2+), and Cd(2+)) by time-resolved optical absorption spectroscopy using the CO flow-flash technique. The time-resolved data were recorded on a ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi049408p
更新日期:2004-12-21 00:00:00