Abstract:
:Two hybrid ribulose-1,5 bisphosphate carboxylase/oxygenase (RubisCO) enzymes were constructed using RubisCO small subunit genes (rbcS) from two eucaryotic marine organisms, Cylindrotheca sp. N1 and Olisthodiscus luteus, cloned downstream of the RubisCO large subunit gene (rbcL) of the cyanobacterium Synechococcus PCC 6301. The expression products synthesized by Escherichia coli JM107 (pVTAC223 and pANOLI) were purified and examined by polyacrylamide gel electrophoresis and compared to the purified products generated by E. coli MV1190 (pBGL710), containing cyanobacterial rbcL and rbcS genes. Both Cylindrotheca and Olisthodiscus small subunits were able to assemble in vivo with the Synechococcus large subunit octamer to form heterologous hexadecameric L8S8 enzymes, the pVTAC223 and pANOLI hybrid enzymes, respectively. Like the Synechococcus RubisCO, the hybrid enzymes were rapidly activated by Mg2+ plus HCO3-, even in the presence of RuBP. The hybrid enzymes, however, were considerably more sensitive to the competitive inhibitor 6-phosphogluconate. Detailed kinetic analysis indicated that while the carboxylase activity of both chimeric enzymes was severely reduced, in the case of the pVTAC223 hybrid enzyme, the degree of partitioning between carboxylation and oxygenation was increased nearly 60% relative to the Synechococcus RubisCO. Other kinetic properties, including the Michaelis constants for the gaseous substrates and RuBP, were altered in the hybrid proteins. These studies also led to the finding that the substrate specificity factor of the Cylindrotheca RubisCO is unusually high.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Read BA,Tabita FRdoi
10.1021/bi00139a018subject
Has Abstractpub_date
1992-06-23 00:00:00pages
5553-60issue
24eissn
0006-2960issn
1520-4995journal_volume
31pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract::The primary electron acceptor pheophytin (Pheo(D1)) plays a crucial role in regulation of forward and backward electron transfer in photosystem II (PSII). It is known that some cyanobacteria control the Pheo(D1) potential in high-light acclimation by exchanging the D1 proteins from different copies of the psbA genes. ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9018829
更新日期:2010-01-26 00:00:00
abstract::The beta-glycosidase from the hyperthermophilic Archaeon Sulfolobus solfataricus hydrolyzes beta-glycosides following a retaining mechanism based upon the action of two amino acids: Glu387, which acts as the nucleophile of the reaction, and Glu206, which acts as the general acid/base catalyst. The activities of inacti...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi981855f
更新日期:1998-12-08 00:00:00
abstract::All CG dinucleotides along exons 5-8 of the p53 tumor suppressor gene contain endogenous 5-methylcytosine (MeC). These same sites (e.g., codons 157, 158, 245, 248, and 273) are mutational hot spots in smoking-induced lung cancer. Several groups used the UvrABC endonuclease incision assay to demonstrate that methylated...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi035259j
更新日期:2004-01-20 00:00:00
abstract::Tumor suppressor Nore1, its acronym coming from novel Ras effector, is one of the 10 members of the Rassf (Ras association domain family) protein family that have been identified. It is expressed as two mRNA splice variants, Nore1A and a shorter isoform, Nore1B. It forms homo- and heterocomplexes through its C-termina...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi3014642
更新日期:2013-02-12 00:00:00
abstract::Despite the expression of only one type of receptor, there is great variation in the ability of different cell types to discriminate between C5a and its more stable metabolite, C5a des Arg74. The mechanism that underlies this phenomenon is not understood but presumably involves differences in the interaction with the ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi990139q
更新日期:1999-07-27 00:00:00
abstract::Leucine aminopeptidase (LAP) is a protease requiring two divalent metal cations per subunit for activity. Zn2+, Mg2+, and Co2+ metal-substituted forms of LAP have been prepared and investigated kinetically. Substitution of metal into the two binding sites independently resulted in the preparation of Zn2+Zn2+, Mg2+Zn2+...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00285a010
更新日期:1983-08-02 00:00:00
abstract::Spectroscopic methods are used to probe the interactions of several anilinoacridine analogues with calf thymus DNA over a wide range of temperatures and sodium chloride concentrations. The structurally similar compounds m-AMSA, AMSA (both active as antitumor agents), and o-AMSA (inactive as an antitumor agent) have be...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00231a025
更新日期:1991-04-30 00:00:00
abstract::Tropomyosins from bovine aorta and pulmonary artery exhibit identical electrophoretic patterns in sodium dodecyl sulfate but differ from tropomyosins of either chicken gizzard or rabbit skeletal muscle. Each of the four tropomyosins binds readily to skeletal muscle F-actin as indicated by their sedimentation with acti...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00299a029
更新日期:1984-02-14 00:00:00
abstract::Immunological methods, in parellel with measurement of ATPase activity, have been used to characterize the reactions of antibodies specific for light chains with myosin and its water-soluble proteolytic subfragments, heavy meromyosin (HMM) and subfragment 1 (HMM S-1). Antiserum to the 5,5'-dithiobis(2-nitro-enzoic aci...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00692a008
更新日期:1975-10-21 00:00:00
abstract::We have studied alkylation of the acetylcholine receptor by N-[3H]ethylmaleimide ([3H]NEM) under various conditions. The radiolabeled preparations were submitted to sodium dodecyl sulfate-polyacrylamide gel electrophoresis to separate the receptor complex into subunits, and the incorporation of 3H into each type of ch...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00306a001
更新日期:1984-05-22 00:00:00
abstract::The structure of human epidermal growth factor (EGF, 53 amino acids) comprises three distinct loops (A, B, and C) connected correspondingly by the three native disulfide bonds, Cys(6)-Cys(20), Cys(14)-Cys(31), and Cys(33)-Cys(42). The connection of Cys(6) and Cys(20) forming the N-terminal A loop is essential for the ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi051399c
更新日期:2005-11-15 00:00:00
abstract::Peptides prepared from the amino termini of pro alpha 1(I) and pro alpha 1(III) collagen chains inhibit the production of pro alpha 1(I) and pro alpha 2 by rat calvaria rna in a reticulocyte cell-free system. The synthesis of other proteins was not altered, suggesting a specific effect on collagen production. Various ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00589a034
更新日期:1979-10-30 00:00:00
abstract::The major carbohydrate fragment from the lipophosphoglycan of Leishmania donovani was generated by mild acid hydrolysis (0.02 N HCl, 5 min, 100 degrees C) and purified by chromatography on DE-52 cellulose and thin layer. By a combination of analyses including gas-liquid chromatography-mass spectrometry and 1H NMR, the...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00393a042
更新日期:1987-09-22 00:00:00
abstract::The mechanism of wild-type and R37A mutant Pseudomonas dacunhae aspartate beta-decarboxylase (ABDC) was studied by rapid-scanning stopped-flow spectrophotometry. Mixing wild-type ABDC with 50 mM disodium l-Asp resulted in the formation of a 325 nm absorption peak within the dead time of the stopped-flow instrument, li...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi100272g
更新日期:2010-06-22 00:00:00
abstract::A pathogenic bacterium has its own mechanisms for not only pathogenic attack but also exogenous invasion defense, in which the bacterial cell wall is the front line of attack and defense. We developed a biochemical lanthanide-encoding approach to quantify the uncanonical d-amino acid (d-X) that was edited in a small p...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.0c00614
更新日期:2020-09-29 00:00:00
abstract::Ribonucleotide reductase from Escherichia coli consists of two nonidentical subunits, named protein B1 (170 000) and protein B2 (87 000). We purified and characterized five monoclonal antibodies against B1 and three against B2 from hybridomas obtained by fusion of spleen cells from immunized mice and the myeloma cell ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00352a018
更新日期:1986-02-25 00:00:00
abstract::The third and most recently identified Parkinson's disease-linked variant of the neuronal protein alpha-synuclein to be identified (E46K) results in widespread brain pathology and early onset Parkinson symptoms (Zarranz et al. (2004) Ann. Neurol. 55, 164-173). Herein, we present biochemical and biophysical characteriz...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi7000246
更新日期:2007-06-19 00:00:00
abstract::Pyrrolidone carboxyl peptidases (PCPs) from hyperthermophiles have a structurally conserved and completely buried Glu192 in the hydrophobic core; in contrast, the corresponding residue in the mesophile protein is a hydrophobic residue, Ile. Does the buried ionizable residue contribute to stabilization or destabilizati...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi052610n
更新日期:2006-06-13 00:00:00
abstract::Proton-detected heteronuclear multiple-bond 1H-13C correlations (HMBC) previously have been used for assignment purposes in a variety of isotopically enriched proteins. In the present study it is demonstrated that the technique yields an almost complete assignment of the natural abundance 13C spectrum of the protein b...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00107a014
更新日期:1991-10-29 00:00:00
abstract::The involvement of calmodulin in the proliferation of Chinese hamster embryo fibroblast cells has been studied with a specific monoclonal antibody to calmodulin. We observed that calmodulin levels increase 2-fold in the late G1 period in these cells, and this coincides with the increase in DNA polymerase alpha activit...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00158a002
更新日期:1992-11-03 00:00:00
abstract::It was previously observed that two steps of the reaction of eukaryotic DNA topoisomerase I (topoisomerization and cleavage) depend upon the conformation of the DNA substrate: in both instances the supercoiled form is a more efficient substrate than the relaxed one. This paper reports the analysis of two other steps o...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00487a024
更新日期:1990-09-04 00:00:00
abstract::The extent of nativeness of the stable conformation of the thermolysin fragment containing the carboxyl-terminal third of the protein (from residues 206 to 316, denoted fragment FII) was examined by its immunogenic and antigenic characteristics. Antisera elicited in rabbits by either intact thermolysin or fragment FII...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00538a007
更新日期:1982-04-27 00:00:00
abstract::R67 dihydrofolate reductase (DHFR) is a novel homotetrameric protein that possesses 222 symmetry and a single, voluminous active site pore. This symmetry poses numerous limitations on catalysis; for example, two dihydrofolate (DHF) molecules or two NADPH molecules, or one substrate plus one cofactor can bind. Only the...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi052504l
更新日期:2006-05-30 00:00:00
abstract::Eukaryotic translation initiation factor 2B (eIF2B) is the guanine nucleotide exchange factor of the GTPase eIF2, which brings the initiator Met-tRNAi to the ribosome in the form of the eIF2-GTP·Met-tRNAi ternary complex (TC). The activity of eIF2B is inhibited by phosphorylation of its substrate eIF2 by several stres...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.7b00957
更新日期:2018-03-06 00:00:00
abstract::Members of the phospholipase C-beta (PLC-beta) family of proteins are activated either by G alpha or G beta gamma subunits of heterotrimeric G proteins. To define specific regions of PLC-beta 3 that are involved in binding and activation by G beta gamma, a series of fragments of PLC-beta 3 as glutathione-S-transferase...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi992021f
更新日期:2000-02-22 00:00:00
abstract::Prolyl endopeptidase (Prep) is a member of the prolyl peptidase family and is of interest because of its unique biochemistry and connections to cognitive function. Using an unbiased mass spectrometry (MS)-based peptidomics platform, we identified Prep-regulated peptides in the central nervous system (CNS) of mice by m...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi901637c
更新日期:2009-12-22 00:00:00
abstract::The Co and Fe K-edge extended X-ray absorption fine structure (EXAFS) spectra of the methionyl aminopeptidase from Escherichia coli (EcMetAP) have been recorded in the presence of 1 and 2 equiv of either Co(II) or Fe(II) (i.e., [Co(II)_(EcMetAP)], [Co(II)Co(II)(EcMetAP)], [Fe(II)_(EcMetAP)], and [Fe(II)Fe(II)(EcMetAP)...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi010837m
更新日期:2001-11-06 00:00:00
abstract::Oxindolealanine-62 lysozyme, formed by reaction with N-bromosuccinimide and purified by using affinity chromatography, was examined in its binding of homologous oligosaccharides of N-acetylglucosamine and in its catalysis of hydrolysis and of transglycosylation of the hexasaccharide of N-acetylglucosamine. Equilibrium...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00558a022
更新日期:1980-08-19 00:00:00
abstract::After protection of cysteine-45 and -82 with iodoacetamide or N-ethylmaleimide, and in the presence of saturating concentrations of substrates, the supernatant isozyme of pig heart aspartate transaminase has been covalently modified at cysteine-390 with 3-bromo-1,1,1-trifluoropropanone. The modified enzyme retains 60-...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00627a004
更新日期:1977-04-19 00:00:00
abstract::Rabbit lung flavin-containing monooxygenase (FMO, EC 1.14.13.8) was denatured, reduced, carboxymethylated, digested with endoproteinase Glu-C or trypsin, and subjected to mass spectrometric analysis. The amino acid sequences of selected peptides were determined by tandem mass spectrometry. Over 90% of rabbit lung FMO ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00105a012
更新日期:1991-10-15 00:00:00