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111.
The light-harvesting pigment-protein complexes allophycocyanin (AP), C-phycocyanin (PC) and phycoerythrocyanin (PEC) of the cyanobacterium Mastigocladus laminosus consist of alpha- and beta-subunits containing about 170 amino-acid residues each. These two subunits form an alpha,beta-monomer, three of which build up a disc-shaped trimer. In this study these phycobiliproteins were crosslinked with bis-imidates. Various spacer lengths of the reagent and various aggregation states of the phycobiliprotein were tested. An intersubunit crosslink could be verified in all three phycobiliproteins. PC-trimers were crosslinked with the homobifunctional reagent dimethyl pimelimidate having a maximal crosslinking distance of 10 A. Two crosslinks could be identified: an intramonomer intersubunit crosslink with a yield of 48% and an intrasubunit crosslink within alpha PC (57%). These products were chemically and enzymatically fragmented and the small crosslinked peptides were isolated and then identified by amino-acid analysis. The following amino acids were crosslinked: alpha-Val 1 with beta-Ala 1 and alpha-Lys 62 with alpha-Lys 134. Both crosslinks could be localized within the known three-dimensional structure of PC. 相似文献
112.
B. Müller E. Schlichting L. Bischoff K. Schügerl 《Applied microbiology and biotechnology》1987,26(1):36-41
Summary Penicillin G was extracted from a model medium with a secondary amine (Amberlite LA-2) as carrier in n-butylacetate as solvent in a 7.6 m high pilot plant Karr-column at different stroke frequencies, throughput of the phases, concentrations of Penicillin G and carrier and ratios of the throughputs of the aqueous and organic phases. Up to penicillin concentrations of 30 gl–1, throughputs of the aqueous phase of 100 lh–1 and throughput ratios of the aqueous phase-to-organic phase of 3, very high degrees of extraction (99%) can be achieved with a penicillin loss below 1%.Symbols a
specific interfacial area with regard to the volume of the continuous phase
- C
partition coefficient
- cA, cA, i
concentration of carrier (sec. amine) in the bulk at the interface
- cAHP, cAHP, i
concentration of complex in the bulk at the interface
- cH
proton concentration
- cHPa, cHPa,i
concentration of free acid in the bulk of the aqueous phase at the interface
- cHPo, cHPo, i
concentration of free acid in the bulk of the organic phase, at the interface
- cP, cP, i
concentration of acid anions in the bulk of the aqueous phase, at the interface
- d32
Sauter droplet diameter
- E
degree of extraction
- f
stroke frequency
- KG
reaction equilibrium constant
- Kphys
distribution coefficient
- N
number of stages in cascade
- t
mean residence time of the aqueous phase
-
aq
throughput of the aqueous phase
-
o
throughput of the organic phase
- Z
dimensionless longitudinal coordinate of the column with regard to its active length (4 m)
-
holdup of the organic phase 相似文献
113.
114.
Subunit II of cytochrome c oxidase from Paracoccus denitrificans. DNA sequence, gene expression and the protein 总被引:14,自引:0,他引:14
P Steinrücke G C Steffens G Panskus G Buse B Ludwig 《European journal of biochemistry》1987,167(3):431-439
Cytochrome c oxidase from the bacterium Paracoccus denitrificans, while being related to the mitochondrial enzyme in many ways, consists of only two to three different subunits. For the identification of its genes, a Paracoccus DNA library was constructed and screened with specific antibodies for expression of cloned inserts in E. coli. A positive clone expressing immunoreactive products in the molecular mass region of authentic subunit II revealed a high homology of its DNA-deduced amino acid sequence with subunit II sequences of the mitochondrial oxidases; several typical features, such as the transmembrane folding pattern and the presumed copper-binding site, are highly conserved between prokaryotic and mitochondrial polypeptides. A comparison with peptide sequencing data of the purified subunit established the presence of a characteristic N-terminal extension as well as a longer C terminus in the initial translation product of the Paracoccus subunit; by mass spectroscopy, the first N-terminally blocked residue of the mature polypeptide was identified as a pyroglutamate. No code abnormalities, but a highly specific codon usage were observed; no evidence for a localization of the subunit I gene directly adjacent to this gene has been obtained. 相似文献
115.
Subcellular location of enzymes involved in the N-glycosylation and processing of asparagine-linked oligosaccharides in Saccharomyces cerevisiae 总被引:3,自引:0,他引:3
A particulate translation system isolated from the yeast Saccharomyces cerevisiae was shown to translate faithfully in-vitro-transcribed mRNA coding for a mating hormone precursor (prepro-alpha-factor mRNA) and to N-glycosylate the primary translation product after its translocation into the lumen of the microsomal vesicles. Glycosylation of its three potential sugar attachment sites was found to be competitively inhibited by acceptor peptides containing the consensus sequence Asn-Xaa-Thr, supporting the view that the glycan chains are N-glycosidically attached to the prepro-alpha-factor polypeptide. The accumulation in the presence of acceptor peptides of a membrane-specific, unglycosylated translation product (pp-alpha-F0) differing in molecular mass from a cytosolically located, protease-K-sensitive alpha-factor polypeptide (pp-alpha-Fcyt) by about 1.3 kDa, suggests that, in contrast to previous reports, a signal sequence is cleaved from the mating hormone precursor on/after translocation. This conclusion is supported by the observation that the multiply glycosylated alpha-factor precursor is cleaved by endoglucosaminidase H to a product with a molecular mass smaller than the primary translation product pp-alpha-Fcyt but larger than the membrane-specific pp-alpha-F0. Translation and glycosylation experiments carried out in the presence of various glycosidase inhibitors (e.g. 1-deoxynojirimycin, N-methyl-1-deoxynojirimyin and 1-deoxymannojirimycin) indicate that the N-linked oligosaccharide chains of the glycosylated prepro-alpha-factor species are extensively processed under the in vitro conditions of translation. From the specificity of the glycosidase inhibitors applied and the differences in the molecular mass of the glycosylated translation products generated in their presence, we conclude that the glycosylation-competent microsomes contain trimming enzymes, most likely glucosidase I, glucosidase II and a trimming mannosidase, which process the prepro-alpha-factor glycans down to the (Man)8(GlcNAc)2 stage. Furthermore, several arguments strongly suggest that these three enzymes, which apparently represent the full array of trimming activities in yeast, are exclusively located in the lumen of microsomal vesicles derived from endoplasmic reticulum membranes. 相似文献
116.
B Kurelec S Britvi? S Krca W E Müller R K Zahn 《Comp. Biochem. Physiol. C, Comp. Pharmacol. Toxicol.》1987,86(1):17-22
Postmitochondrial fractions from marine sponges Geodia cydonium, Tethya aurantium, Verongia aerophoba and Pellina semitubulosa activate precarcinogenic aromatic amine 2-aminoanthracene, but not precarcinogenic polycyclic aromatic hydrocarbon benzo(a)pyrene, to Salmonella typhimurium TA 98 mutagens. All four sponge species lack a benzo(a)pyrene monooxygenase activity, but possesses the enzyme activity whose characteristics (selective activation of aromatic amines, NADPH-dependency, pH optimum at 8.4) are similar to FAD-containing monooxygenase. Tethya postmitochondrial fraction possesses an UDP-glucuronyl transferase activity which catalyzes the conjugation of a considerable part of metabolized 2-acetylamino [9-14C]fluorene to water soluble glucuronides. The possible ecological significance of exuded aromatic amine metabolites as well as the significance of the presence of the selective potential for the activation of aromatic amines to mutagens among sponges for our understanding of the fate and effects of carcinogens in the marine environment are discussed. 相似文献
117.
Summary The receptor lymph of campaniform sensilla on the halteres of the blowfly, Calliphora vicina, was analyzed histochemically. Acid mucopolysaccharides were demonstrated by a test for iron-binding capacity (Hale-reaction). Further characterization by enzyme treatment showed that the receptor lymph contains hyaluronic acid and/or chondroitin sulfate. Ultrahistochemical studies gave evidence for glycoproteins in the inner and outer receptor lymph space. The significance of acid mucopolysaccharides for arthropod sensilla is discussed. 相似文献
118.
Hydrogen-deuterium exchange is measured for the buried primary amide groups of Asn-43 and Asn-44 in bovine pancreatic trypsin inhibitor. Amide protons trans and cis to the amide carbonyl oxygen (HE and HZ, respectively) exchange at indistinguishable rates. Uncorrelated exchange of HE and HZ is established for both residues by following the nuclear Overhauser enhancement from HE to HZ during the deuterium exchange. The exchange of Asn-43 and Asn-44 side-chain protons differs qualitatively from exchange of primary amide groups in fully solvated model compounds, for which HE generally exchanges faster than HZ. The equal rates for the buried primary amide HE and HZ in BPTI are not a consequence of coupled exchange. The data indicate rapid rotation around the CO-NH2 bond for both Asn-43 and Asn-44 and suggest considerable lability of intramolecular hydrogen bonds. The side chain of Asn-43 has all of its polar atoms integrated into the very stable hydrogen-bonded structure of the protein. Asn-44 is hydrogen-bonded to side chains and to a buried water molecule. Solvent isotope exchange is several orders of magnitude more restricted by protein secondary and tertiary structure than the CO-NH2 rotation, indicating that N delta H2 groups flip many times before hydrogen isotope exchange occurs. 相似文献
119.
Lipopeptides of the N-terminus of Escherichia coli lipoprotein: synthesis, mitogenicity and properties in monolayer experiments 总被引:3,自引:0,他引:3
W Prass H Ringsdorf W Bessler K H Wiesmüller G Jung 《Biochimica et biophysica acta》1987,900(1):116-128
The N-terminal part of the lipoprotein from the outer membrane of Escherichia coli, tripalmitoyl-S-glyceryl-L-Cys-Ser and analogs with longer sequences, are polyclonal activators for B-lymphocytes. Triple-chain lipopeptides also constitute efficient low-molecular-weight carrier/adjuvant systems, which can be linked to antigens to yield immunogens for antibody production without further additives. This is the first report of monolayer experiments with chemically well defined, synthetic lipopeptide mitogens with the composition of the N-terminus of an important bacterial membrane protein. Various derivatives of the lipoprotein N-terminus were synthesized. These lipopeptides differed in the length of the peptide moiety, the number of fatty acid residues, and protective groups. In order to obtain the surface areas for the lipopeptides in isotherms and hysteresis isotherms, monolayer experiments with a computer-controlled film balance were performed. To get some information about the interaction of these compounds with typical membrane lipids mixed monolayers were formed from triple-chain lipopeptides with dipalmitoylphosphatidylcholine and cholesterol. A comparison of the mitogenic response of the compounds was made in an in vitro system with B-lymphocytes from Balb/c mice. 相似文献
120.
Structural elements of bactopterin from Pseudomonas carboxydoflava carbon monoxide dehydrogenase 总被引:2,自引:0,他引:2
Bactopterin is a novel pterin occurring in bacterial molybdoenzymes as the organic portion of the molybdenum cofactor. Its structure is investigated here. The compound contains a single pterin ring and carries a side chain at carbon atom 6 of the pterin nucleus as indicated by the formation of pterin-6-carboxylic acid upon alkaline permanganate oxidation. Studies with phosphate-cleaving enzymes revealed the presence of two monophosphoric acid monoesters. The affinity of reduced bactopterin for thiol-Sepharose points to the presence of thiol(s) in active bactopterin. 相似文献