Abstract:
:The 600-MHz proton spectrum of dethiobiotin (prepared from d-biotin with Raney nickel) was measured in order to gain information pertaining to its stereochemical homogeneity. The spectrum demonstrated clearly that the material is a 6:1 mixture of two stereoisomers. The cis compound, corresponding to the stereochemistry of d-biotin, is the major isomer. Two biotinyl- and two dethiobiotinylinsulins were prepared in which the distance between the biotins and insulin was varied by interposition of spacer arms. The synthesis of these compounds involved repeated N-hydroxysuccinimido ester condensations. Biotin N-hydroxysuccinimido ester, dethiobiotin N-hydroxysuccinimido ester, 6-aminohexanoic acid, and N-[3-[(3-aminopropyl)carboxyamino]-propyl]succinamic acid N-tert-butyl ester served as the building blocks for the spacers. The latter compound was prepared from N-[3-[(3-aminopropyl)amino]propyl]succinamic acid sulfate by the use of a selective amino-protecting method based on the differential stability toward acid of citraconyl and tert-butoxycarbonyl amino-protecting groups. The structure of N-[3-[(3-aminopropyl)amino]propyl]succinamic acid sulfate was established unequivocally by X-ray diffraction. The attachment of the biotinylated spacers to the insulin was exclusively at the N alpha, B1 position. Homogeneity of the final products as well as of the intermediates used in their synthesis was established by thin-layer chromatography, by high-pressure liquid chromatography, and in most instances by elemental analysis. The ratio of 6-aminohexanoic acid to lysine in hydrolysates of the insulin derivatives was in agreement with theory. The insulin derivatives were required for a study on the effect of avidin on their ability to interact with insulin receptors on rat epididymal adipocytes, which is described in the accompanying paper.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Hofmann K,Zhang WJ,Romovacek H,Finn FM,Bothner-By AA,Mishra PKdoi
10.1021/bi00307a002subject
Has Abstractpub_date
1984-06-05 00:00:00pages
2547-53issue
12eissn
0006-2960issn
1520-4995journal_volume
23pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract::The effects of detergent [deoxycholate (DOC) and phospholipid [dimyristoylphosphatidylcholine (DMPC)] environments on the rotational dynamics of the single tryptophan residue 26 of bacteriophage M13 coat protein have been investigated by using time-resolved single photon counting measurements of the fluorescence inten...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00441a035
更新日期:1989-07-25 00:00:00
abstract::The single reactive sulfhydryl group, located in the active site of each subunit of dimeric creatine kinase from rabbit muscle (isozyme MM), was selectively labeled with 3-(4-maleimidylphenyl)-7-(diethylamino)-4-methylcoumarin (CPM). Isozyme BB, purified to homogeneity from rabbit brain, was conjugated with the sulfhy...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00292a018
更新日期:1983-11-08 00:00:00
abstract::The systems of tRNA glycylation belong to the most complex aminoacylation systems since neither the oligomeric structure of glycyl-tRNA synthetases (GlyRS) nor the discriminator bases in tRNAGly are conserved in the phylae. To better understand the structure-function relationship in glycylation systems of various orig...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi991392t
更新日期:1999-10-05 00:00:00
abstract::The X-ray crystal structure of the At5g18200.1 protein has been determined to a nominal resolution of 2.30 A. The structure has a histidine triad (HIT)-like fold containing two distinct HIT-like motifs. The sequence of At5g18200.1 indicates a distant family relationship to the Escherichia coli galactose-1-P uridylyltr...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi052232m
更新日期:2006-03-14 00:00:00
abstract::The chemical shifts of all the aromatic proton and anomeric proton resonances of NADP+, NADPH, and several structural analogues have been determined in their complexes with Lactobacillus casei dihydrofolate reductase by double-resonance (saturation transfer) experiments. The binding of NADP+ to the enzyme leads to lar...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00557a015
更新日期:1980-08-05 00:00:00
abstract::Molecular dynamics (MD) simulations of Thermus thermophilus chorismate mutase substrate complex (TtCM x S) have been carried out at 298 K, 333 K, and the temperature of optimum activity: 343 K. The enzyme exists as trimeric subunits with active sites shared between two neighboring subunits. Two features distinguish in...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0604227
更新日期:2006-07-18 00:00:00
abstract::The alpha-helix [Pauling, L., Corey, R. B., & Branson, H. R. (1951) Proc. Natl. Acad. Sci. U.S.A. 37, 205-211] is a common motif in both proteins and peptides. Despite intense investigation, predictive understanding of helices is still lacking. A recent hypothesis [Presta, L. G., & Rose, G. D. (1988) Science 240, 1632...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00081a001
更新日期:1993-08-03 00:00:00
abstract::To develop molecules capable of directly probing the catechol binding region of the beta(2)-adrenergic receptor (beta(2)AR), novel benzophenone- and fluorenone-based beta(2)AR antagonists were prepared as potential photoaffinity probes. While the benzophenone-containing ligands bound with relatively modest affinity, o...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi001342k
更新日期:2000-10-24 00:00:00
abstract::The role of histidine in catalysis by prostaglandin H synthase has been investigated using chemical modification with diethyl pyrocarbonate (DEPC), an agent that has been found to rather selectively derivatize histidine residues in proteins under mild conditions. Incubation of the synthase apoprotein with DEPC at pH 7...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00124a013
更新日期:1992-03-10 00:00:00
abstract::The PLL(PTE-like lactonase)-group of enzymes within the amidohydrolase superfamily hydrolyze N-acyl-homoserine lactones (AHLs) that are involved in bacterial quorum-sensing pathways. These enzymes possess the (beta/alpha)(8)-barrel fold and serve as attractive templates for in vitro evolution and engineering of quorum...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9004045
更新日期:2009-05-26 00:00:00
abstract::Regulation of adenylate cyclase by alpha 2 adrenergic receptors requires the inhibitory guanine nucleotide binding protein Ni. A role for this protein has also been suggested in the high-affinity binding of agonists to the alpha 2 receptor. We recently reported that alkaline treatment can selectively inactivate alpha ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00386a061
更新日期:1987-06-16 00:00:00
abstract::Western blot analysis, using a polyclonal antibody to the 240-kDa endogenous inhibitor of the 20 S proteasome, revealed that the inhibitor is a component of the 26 S complex. Although isolated inhibitor displayed a single 40-kDa band on SDS-PAGE, the antibody detected a 55-kDa component in the 26 S proteasome complex....
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00163a001
更新日期:1992-12-08 00:00:00
abstract::The DNA binding domain (DBD) of p53 folds by a complex mechanism that involves parallel pathways and multiple intermediates, both on- and off-pathway. This heterogeneity renders DBD particularly susceptible to misfolding and aggregation. The origins of parallel folding mechanisms are not well understood. DBD folding h...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi062106y
更新日期:2007-03-13 00:00:00
abstract::The targeting of the insulin-responsive glucose transporter, GLUT-4, to an intracellular compartment in adipocytes and muscle is one of the key features responsible for the unique insulin sensitivity of this transporter. Through expression of epitope-tagged GLUT-4 mutants in 3T3-L1 adipocytes, two motifs have been ide...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi980988y
更新日期:1999-02-02 00:00:00
abstract::The iron-chelating peptide vibriobactin of the pathogenic Vibrio cholerae is assembled by a four-subunit nonribosomal peptide synthetase complex, VibE, VibB, VibH, and VibF, using 2,3-dihydroxybenzoate and L-threonine as precursors to two 2,3-dihydroxyphenyl- (DHP-) methyloxazolinyl groups in amide linkage on a norspe...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi010937s
更新日期:2001-09-04 00:00:00
abstract::Steady-state kinetic parameters were determined for the human leukocyte elastase catalyzed hydrolysis of a series of peptide-based thiobenzyl esters and p-nitroanilides. The peptide units are MeOSuc-Val, MeOSuc-Alan-Pro-Val (n = 0-2), and MeOSuc-Alan-Pro-Ala (n = 1 or 2). The results of this study suggest five importa...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00379a015
更新日期:1987-03-10 00:00:00
abstract::The lipase from Staphylococcus hyicus (SHL) displays a high phospholipase activity whereas the homologous S. aureus lipase (SAL) is not active or hardly active on phospholipid substrates. Previously, it has been shown that elements within the region comprising residues 254-358 are essential for the recognition of phos...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi990096d
更新日期:1999-07-20 00:00:00
abstract::The interaction of nile red (NR) with apomyoglobin (ApoMb) in the native (pH 7) and molten globule (pH 4) states was investigated using experimental and computational methods. NR binds to hydrophobic locations in ApoMb with higher affinity (K(d) = 25 +/- 5 microM) in the native state than in the molten globule state (...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi051905y
更新日期:2006-04-25 00:00:00
abstract::The effect of monoclonal anti-(DNA polymerase alpha) immunoglobulin G (IgG) and F(ab) fragments on DNA replication in lysolecithin-permeabilized human cells and on DNA polymerase alpha activity was determined. DNA polymerase alpha activity in vitro was inhibited equally by the same concentrations of monoclonal IgGs an...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00346a061
更新日期:1985-12-03 00:00:00
abstract::The light harvesting 1 antenna (LH1) complex from Rhodobacter sphaeroides funnels excitation energy to the photosynthetic reaction center. Our ultimate goal is to build up the structure of LH1 from structures of its individual subunits, much as the antenna can self-assemble from its components in membrane-mimicking de...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi011576j
更新日期:2002-01-08 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::We have determined the nucleotide sequence and the derived amino acid sequence of the phosphoprotein-encoding region of the chicken vitellogenin III gene. The sequence of this minor vitellogenin could be aligned with exon 22 up to exon 27 of the previously sequenced major vitellogenin II gene (van het Schip et al., 19...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00432a034
更新日期:1989-03-21 00:00:00
abstract::The crystal structure of the complex between the quinoprotein methylamine dehydrogenase (MADH) and the type I blue copper protein amicyanin, both from Paracoccus denitrificans, has been determined at 2.5-A resolution using molecular replacement. The search model was MADH from Thiobacillus versutus. The amicyanin could...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00136a006
更新日期:1992-06-02 00:00:00
abstract::Differential scanning calorimetry and CD spectrometry were employed to study the thermal unfolding of light meromyosin (LMM) prepared from carp acclimated to different temperatures. The transition temperatures given by the major peaks at pH 8.0 in 0.6 M KCl for LMM from carp acclimated to 10 degrees C were 32.5 and 39...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9701181
更新日期:1997-07-29 00:00:00
abstract::From the larger set of 191 variants at all the variable contact positions in the turkey ovomucoid third domain, we selected a subset that consists of Asp, Glu, His, and Lys residues at eight of the nine contiguous P6-P3' positions (residues 13-21), the exception being P3-Cys16 which is involved in a conserved disulfid...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0269512
更新日期:2003-03-18 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::A wide range of two-dimensional 1H NMR experiments have been used to completely assign the 500-MHz 1H NMR spectrum of recombinant Ca2+-saturated bovine calbindin D9k (76 amino acids, Mr = 8500). In solution, calbindin D9k exists as an equilibrium mixture of isoforms with trans (75%) and cis (25%) isomers of the peptid...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00443a043
更新日期:1989-08-22 00:00:00
abstract::Protein disulfide isomerase (PDI) utilizes the active site sequence Cys-Gly-His-Cys (CGHC; E degrees ' = -180 mV) to effect thiol-disulfide interchange during oxidative protein folding. Here, the Cys-Gly-Cys-NH(2) (CGC) peptide is shown to have a disulfide reduction potential (E degrees ' = -167 mV) that is close to t...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi026993q
更新日期:2003-05-13 00:00:00
abstract::The interactions of HIV-1 Env (gp120-gp41) with CD4 and coreceptors trigger a barrage of conformational changes in Env that drive the membrane fusion process. Various regions of gp41 have profound effects on HIV entry and budding. However, the precise interactions between gp41 and the membrane have not been elucidated...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi701920f
更新日期:2008-02-19 00:00:00
abstract::The soluble domains of the b and b' subunits of the ATP synthase of the cyanobacterium Synechocystis PCC 6803 were expressed with His tags attached to their N-termini. Following purification, the polypeptides were characterized by chemical cross-linking, analytical ultracentrifugation, and circular dichroism spectrosc...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi001821j
更新日期:2001-01-09 00:00:00