Abstract:
:The solution structure and dynamics of human insulin are investigated by 2D 1H NMR spectroscopy in reference to a previously analyzed analogue, des-pentapeptide(B26-B30) insulin (DPI; Hua, Q.X., & Weiss, M.A. (1990) Biochemistry 29, 10545-10555). This spectroscopic comparison is of interest since (i) the structure of the C-terminal region of the B-chain has not been determined in the monomeric state and (ii) the role of this region in binding to the insulin receptor has been the subject of long-standing speculation. The present NMR studies are conducted in the presence of an organic cosolvent (20% acetic acid), under which conditions both proteins are monomeric and stably folded. Complete sequential assignment of human insulin is obtained and leads to the following conclusions. (1) The secondary structure of the insulin monomer (three alpha-helices and B-chain beta-turn) is similar to that observed in the 2-Zn crystal state. (2) The folding of DPI is essentially the same as the corresponding portion of intact insulin, in accord with the similarities between their respective crystal structures. However, differences between insulin and DPI are observed in the extent of conformational broadening of amide resonances, indicating that the presence or absence of residues B26-B30 influences the overall dynamics of the protein on the millisecond time scale. (3) Residues B24-B28 adopt an extended configuration in the monomer and pack against the hydrophobic core as in crystallographic dimers; residues B29 and B30 are largely disordered. This configuration differs from that described in a more organic milieu (35% acetonitrile; Kline, A.D., & Justice, R.M., Jr. (1990) Biochemistry 29, 2906-2913), suggesting that the conformation of insulin in the latter study may have been influenced by solvent composition. (4) The insulin fold is shown to provide a model for collective motions in a protein with implications for the mechanism of protein-protein recognition. To our knowledge, this paper describes the first detailed analysis of a protein NMR spectrum under conditions of extensive conformational broadening. Such an analysis is made possible in the present case by comparative study of an analogue (DPI) with more tractable spectroscopic properties.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Hua QX,Weiss MAdoi
10.1021/bi00236a025subject
Has Abstractpub_date
1991-06-04 00:00:00pages
5505-15issue
22eissn
0006-2960issn
1520-4995journal_volume
30pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract::The activity of serine carboxypeptidases is dependent on a catalytic triad, an oxyanion hole, and a binding site equivalent to those found in the serine endopeptidases. The action of carboxypeptidase Y on substrates containing amino acids, alcohols, and amines as leaving groups is described. It is demonstrated that th...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi952758e
更新日期:1996-06-04 00:00:00
abstract::Chlorophyll a fluorescence emission is widely used as a noninvasive measure of a number of parameters related to photosynthetic efficiency in oxygenic photosynthetic organisms. The most important component for the estimation of photochemistry is the relative increase in fluorescence yield between dark-adapted samples ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9806854
更新日期:1998-08-04 00:00:00
abstract::The ligation state of hemoglobin during its cross-linking by glycolaldehyde influences the ultimate oxygen affinity of the cross-linked protein. Thus, if the cross-linking is performed with carbonmonoxy-hemoglobin, the oxygen affinity increases slightly to a P50 of 7 mmHg from a P50 of 9 mmHg for unmodified hemoglobin...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00417a064
更新日期:1988-08-23 00:00:00
abstract::Aminoimidazole riobnucleotide carboxylase, the sixth step in the purine biosynthetic pathway, catalyzes the conversion of aminoimidazole ribonucleotide (AIR) to carboxyaminoimidazole ribonucleotide (CAIR). The gene products of the purE and purK genes (PurE and PurK, respectively) thought to be responsible for this act...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00136a016
更新日期:1992-06-02 00:00:00
abstract::Flash-induced Fourier transform infrared (FTIR) difference spectra for the four-step S-state cycle and the effects of global (15)N- and (13)C-isotope labeling on the difference spectra were examined for the first time in the mid- to low-frequency (1200-800 cm(-1)) as well as the mid-frequency (1700-1200 cm(-1)) region...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0362323
更新日期:2004-06-15 00:00:00
abstract::Spectrophotometric titration of pyridoxamine-pyruvate transaminase (EC 2.6.1.30) with pyridoxal at pH 7.15 gives four equivalent binding sites per tetramer. The pH dependence of the equilibrium constant for the association of 5'-deoxypyridoxal with the active site lysine residue was determined spectrophotometrically. ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00643a013
更新日期:1977-11-29 00:00:00
abstract::The lateral and rotational dynamics of phosphoinositides and their interactions with proteins were characterized using pyrene-labeled lipid analogues. In these systems, the collision frequency of pyrene-labeled lipids was studied by monitoring the monomeric pyrene fluorescence yield as a function of their mole fractio...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00028a022
更新日期:1995-07-18 00:00:00
abstract::A variety of chronic inflammatory conditions are associated with an increased risk for the development of cancer. Because of the numerous links between DNA oxidative damage and carcinogenesis, a potential role for leukocyte-generated oxidants in these processes has been suggested. In the present study, we demonstrate ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi992809y
更新日期:2000-05-09 00:00:00
abstract::The binding of Ca2+ and the resulting change in catalytic specificity that allows phosphorylation of the calcium ATPase of sarcoplasmic reticulum by ATP were examined by measuring the amount of phosphoenzyme formation from [32P]ATP, or 45Ca incorporation into vesicles, after the simultaneous addition of ATP and EGTA a...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00423a018
更新日期:1988-11-15 00:00:00
abstract::The removal of damaged bases by DNA glycosylases is thought to be effectively irreversible, because of an overall equilibrium that favors hydrolysis over synthesis of the N-glycosyl bond. Surprisingly, human alkyladenine DNA glycosylase (AAG) can make damaged DNA by catalyzing formation of an N-glycosyl bond between 1...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi101380d
更新日期:2010-10-26 00:00:00
abstract::Carbonic anhydrase (CA) catalyzes the reversible hydration of carbon dioxide to hydrogen carbonate, and its role in maintaining pH balance has made it an attractive drug target. Steroidal sulfamate esters, inhibitors of the cancer drug target steroid sulfatase (STS), are sequestered in vivo by CA II in red blood cells...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi902178w
更新日期:2010-04-27 00:00:00
abstract::The growth and shortening of microtubules in guanosine triphosphate-(GTP-) mediated dynamic instability has previously been observed to occur at rates which are remarkably variable (Gildersleeve et al., 1992, Chrétien et al., 1995). Neighboring microtubules observed simultaneously can grow or shorten at different rate...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi961070e
更新日期:1996-11-12 00:00:00
abstract::The presence of high titers of anti-cardiolipin antibodies (ACA's) of autoimmune origin, which are known to bind to plasma beta2-glycoprotein I (aka apolipoprotein H), correlates clinically with autoimmune recurrent thrombosis. Soluble beta2-glycoprotein I binds to solid-phase ACA (immobilized on a surface plasmon res...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9807207
更新日期:1998-11-17 00:00:00
abstract::The complete amino acid sequence was determined for bovine ubiquitin, and adenylate cyclase stimulating polypeptide, which is probably represented universally in living cells. Ubiquitin has a molecular weight of 8451 and consists of a single polypeptide chain containing 74 amino acid residues. It contains four argini...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00681a026
更新日期:1975-05-20 00:00:00
abstract::In Sphingomonas CHY-1, a single ring-hydroxylating dioxygenase is responsible for the initial attack of a range of polycyclic aromatic hydrocarbons (PAHs) composed of up to five rings. The components of this enzyme were separately purified and characterized. The oxygenase component (ht-PhnI) was shown to contain one R...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0611311
更新日期:2006-10-10 00:00:00
abstract::The relationship between triacylglycerol and monoacylglycerol hydrolyzing activities of purified rat heart lipoprotein lipase was studied using emulsified trioleoylglycerol and micellar or albumin-bound monooleoylglycerol as substrates. The maximal reaction rates obtained with the two substrates were similar (650 and ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00654a017
更新日期:1976-05-04 00:00:00
abstract::Amyloid beta peptide (A beta) is released into the media of a variety of cells in culture during normal metabolism. The discovery of several missense mutations within or flanking the A beta region of the beta amyloid precursor protein (beta APP) in familial Alzheimer's disease provides strong evidence for a role of al...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00181a016
更新日期:1994-04-19 00:00:00
abstract::The proton-translocating NADH-quinone oxidoreductase (NDH-1) of Paracoccus denitrificans is composed of 14 different subunits (Nqo1-Nqo14). Of these, seven subunits (Nqo7, Nqo8, and Nqo10-14) which are equivalent to the mitochondrial DNA-encoded subunits of complex I constitute the membrane segment of the enzyme compl...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi034166z
更新日期:2003-04-22 00:00:00
abstract::Protein folding is a process in which an extended polypeptide chain acquires compact packing through the formation of specific secondary and tertiary structures and hydrophobic interactions. Although much attention has been paid to secondary and tertiary structures, there is no definitive view about the relationship b...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00182a019
更新日期:1994-04-26 00:00:00
abstract::The nucleotide binding site of the uncoupling protein (UCP) from brown adipose tissue was mapped by photoaffinity labeling with 2-azidoadenosine 5'-triphosphate (2-azido-ATP) and by affinity labeling with 3'-O-(5-fluoro-2,4-dinitrophenyl)adenosine 5'-triphosphate (FDNP-ATP). Both analogs bind with high affinity and sp...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00158a017
更新日期:1992-11-03 00:00:00
abstract::A recently reported variant of human transthyretin (TTR), Leu-55-Pro, implicated as the causative agent in early-onset familial amyloid polyneuropathy was expressed and characterized, and its denaturation pathway and amyloidogenicity were compared to those of wild-type transthyretin. The overlap-extension polymerase c...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00096a024
更新日期:1993-11-16 00:00:00
abstract::Dopamine (DA) is an unstable neurotransmitter that readily oxidizes to the DA quinone and forms reactive oxygen species, such as superoxide and hydrogen peroxide. The oxidized dopamine also forms thiol conjugates with sulfhydryl groups on cysteine, glutathione, and proteins. In the present study, we determined the red...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0614434
更新日期:2006-12-19 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::A gentle and rapid isolation procedure is described yielding fractions containing better than 95% pure OmpF porin of Escherichia coli BE with different amounts of bound lipopolysaccharide (LPS). The procedure employs continuous free-flow electrophoresis (FFE) in the presence of detergent above its critical micelle con...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00436a010
更新日期:1989-05-16 00:00:00
abstract::The production of proteins via recombinant DNA technology often requires the in vitro folding of inclusion bodies, which are protein aggregates. To create a more efficient redox buffer for the in vitro folding of disulfide containing proteins, aromatic thiols were investigated for their ability to increase the folding...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi034305c
更新日期:2003-10-14 00:00:00
abstract::Much interest is currently focused on understanding the detailed contribution that particular amino acid residues make in protein structure and function. Although the use of site-directed mutagenesis has greatly contributed to this goal, the approach is limited to the standard repertoire of twenty amino acids. Fluorin...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9617973
更新日期:1997-03-18 00:00:00
abstract::Ligand-dependent site-site (or subunit-subunit) interactions provide the basis for explaining cooperativity in chemical reactions. Even in the simplest possible nonaggregating system, interpretation of the interactions in terms of structural details requires an explicit assumption (or model) for the binding of the lig...
journal_title:Biochemistry
pub_type: 杂志文章,评审
doi:10.1021/bi00106a004
更新日期:1991-10-22 00:00:00
abstract::A general protocol for preparation of oligonucleotides containing intrastrand cross-links between the exocyclic amino groups of adjacent deoxyguanosines has been developed. A series of 2, 3, and 4 methylene cross-links was incorporated site-specifically into an 11-mer (5'-GGCAGGTGGTG-3', cross-linked positions are und...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi010450j
更新日期:2002-03-05 00:00:00
abstract::Mitochondria act as a focal point for upstream apoptosis signals by releasing cytochrome c into the cytosol, leading to the activation of caspases and subsequent cell death. Members of the Bcl-2 protein family regulate this phenomenon by heterodimerization via the BH3 domain of proapoptotic members opposing their pro-...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0110286
更新日期:2002-07-09 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