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1.
Phys Rev Lett ; 92(7): 071102, 2004 Feb 20.
Article in English | MEDLINE | ID: mdl-14995836

ABSTRACT

We present the results of a search for point sources of high-energy neutrinos in the northern hemisphere using AMANDA-II data collected in the year 2000. Included are flux limits on several active-galactic-nuclei blazars, microquasars, magnetars, and other candidate neutrino sources. A search for excesses above a random background of cosmic-ray-induced atmospheric neutrinos and misreconstructed downgoing cosmic-ray muons reveals no statistically significant neutrino point sources. We show that AMANDA-II has achieved the sensitivity required to probe known TeV gamma-ray sources such as the blazar Markarian 501 in its 1997 flaring state at a level where neutrino and gamma-ray fluxes are equal.

2.
Phys Rev Lett ; 90(25 Pt 1): 251101, 2003 Jun 27.
Article in English | MEDLINE | ID: mdl-12857122

ABSTRACT

Data from the AMANDA-B10 detector taken during the austral winter of 1997 have been searched for a diffuse flux of high energy extraterrestrial muon neutrinos. This search yielded no excess events above those expected from background atmospheric neutrinos, leading to upper limits on the extraterrestrial neutrino flux measured at the earth. For an assumed E-2 spectrum, a 90% classical confidence level upper limit has been placed at a level E2Phi(E)=8.4 x 10(-7) cm(-2) s(-1) sr(-1) GeV (for a predominant neutrino energy range 6-1000 TeV), which is the most restrictive bound placed by any neutrino detector. Some specific predicted model spectra are excluded. Interpreting these limits in terms of the flux from a cosmological distributions of sources requires the incorporation of neutrino oscillations, typically weakening the limits by a factor of 2.

3.
Nature ; 410(6827): 441-3, 2001 Mar 22.
Article in English | MEDLINE | ID: mdl-11260705

ABSTRACT

Neutrinos are elementary particles that carry no electric charge and have little mass. As they interact only weakly with other particles, they can penetrate enormous amounts of matter, and therefore have the potential to directly convey astrophysical information from the edge of the Universe and from deep inside the most cataclysmic high-energy regions. The neutrino's great penetrating power, however, also makes this particle difficult to detect. Underground detectors have observed low-energy neutrinos from the Sun and a nearby supernova, as well as neutrinos generated in the Earth's atmosphere. But the very low fluxes of high-energy neutrinos from cosmic sources can be observed only by much larger, expandable detectors in, for example, deep water or ice. Here we report the detection of upwardly propagating atmospheric neutrinos by the ice-based Antarctic muon and neutrino detector array (AMANDA). These results establish a technology with which to build a kilometre-scale neutrino observatory necessary for astrophysical observations.

4.
Mol Biochem Parasitol ; 95(1): 53-68, 1998 Sep 01.
Article in English | MEDLINE | ID: mdl-9763289

ABSTRACT

The protozoal parasite Trypanosoma brucei depends on a mitochondrial non-cytochrome terminal oxidase known as the trypanosome alternative oxidase (TAO) in its mammalian host. We have recently cloned the cDNA from T. brucei bloodstream form and have characterized a 33 kDa mitochondrial protein as TAO. Here we report that the TAO is a single copy gene in T. brucei and its expression is down regulated at the level of transcript abundance during differentiation from the bloodstream to the procyclic trypanosomes. Like other alternative oxidases (AOXs) cloned from different plants and fungi, TAO possesses the conserved sequences at the centrally located predicted membrane spanning domains and the signature sequence at the C-terminal hydrophilic domain for a pair of putative iron binding motifs (E-X-X-H). Phylogenetic analysis of the deduced protein sequences of eight different alternative oxidases cloned from different plants and fungi revealed that TAO is more closely related to the alternative oxidases of the fungi clade than that of plants. TAO has been functionally expressed in Escherichia coli. In the first of the two putative iron binding motifs, site-directed mutagenesis of E215 to A, L, N and Q resulted in the loss of the ability of the TAO gene to complement the heme deficiency of the E. coli mutants (SASX41B and GE1387) by conferring on them a CN-insensitive pathway of respiration. The conservative substitution of E215 by aspartate and histidine reduced the growth of the E. coli auxotrophs by approximately 80%. The mutations apparently did not have any effect on the stability of the expressed protein as revealed by the immunoblot analysis of the bacterial protein using TAO monoclonal antibody, which we have developed. Together, these points suggest that E215 plays an important role in the function of TAO. The steady state level of TAO mRNA is down-regulated in the procyclic stage presumably accounting for the low levels of TAO protein in these forms.


Subject(s)
Oxidoreductases/genetics , Oxidoreductases/metabolism , Trypanosoma brucei brucei/enzymology , Aldehyde Oxidoreductases/genetics , Aldehyde Oxidoreductases/metabolism , Amino Acid Sequence , Animals , Antibodies, Monoclonal , Antigens, Protozoan/immunology , Blotting, Northern , Blotting, Southern , DNA, Complementary , Electrophoresis, Polyacrylamide Gel , Escherichia coli/genetics , Gene Expression Regulation, Enzymologic , Genetic Complementation Test , Immunoblotting , Life Cycle Stages , Mitochondrial Proteins , Molecular Sequence Data , Mutagenesis , Oxidoreductases/chemistry , Phylogeny , Plant Proteins , Plasmids , Trypanosoma brucei brucei/genetics , Trypanosoma brucei brucei/growth & development
6.
J Eukaryot Microbiol ; 42(5): 467-72, 1995.
Article in English | MEDLINE | ID: mdl-7581322

ABSTRACT

The glycerophosphate oxidase (GPO), the unique terminal oxidase of bloodstream trypanosome (TAO), appears to be functionally similar to the alternative oxidases of some plants and higher fungi. Immunoblotting of mitochondrial proteins of bloodstream trypomastigotes of Trypanosoma brucei with monoclonal or polyclonal antibodies to Sauromatum guttatum (voodoo lily) and Symplocarpus foetidus (skunk cabbage) alternative oxidases respectively revealed two proteins of about 33 kDa (p33) and 68 kDa (p68). These proteins are not present in procyclic trypomastigotes. Electrophoresis under rigorous denaturing conditions indicated p68 to be the dimer of p33. Indirect immunofluorescent studies of bloodstream and procyclic trypomastigotes with monoclonal antibody to plant alternative oxidase also showed the localization of 33 kDa protein in the mitochondria of the bloodstream trypomastigotes. The functional TAO activity could be solubilized efficiently from the mitochondrial membrane of the bloodstream trypomastigotes by 1% NP-40 or 10 mM lauryl maltoside. When fractionated by Superose 12 gel filtration chromatography, p33 was co-purified with the TAO enzymatic activity. The apparent molecular size of the active enzyme complex was found to be 160 kDa. Gradual disappearance of the 33 kDa protein and the TAO enzymatic activity were well correlated during in vitro differentiation of the bloodstream to procyclic trypomastigotes. This study implies that the net biosynthesis of p33, an essential subunit of TAO, is decreased during differentiation from bloodstream to procyclic trypomastigotes.


Subject(s)
Mitochondria/enzymology , Oxidoreductases/isolation & purification , Trypanosoma brucei brucei/enzymology , Animals , Antibodies, Monoclonal , Cross Reactions , Mitochondrial Proteins , Molecular Weight , Oxidoreductases/antagonists & inhibitors , Oxidoreductases/chemistry , Oxidoreductases/immunology , Oxidoreductases/metabolism , Plant Proteins , Plants/enzymology , Plants/immunology , Potassium Cyanide/pharmacology , Salicylamides/pharmacology , Trypanocidal Agents/pharmacology , Trypanosoma brucei brucei/drug effects , Trypanosoma brucei brucei/growth & development
7.
J Pharm Sci ; 84(4): 448-55, 1995 Apr.
Article in English | MEDLINE | ID: mdl-7629735

ABSTRACT

X-ray diffraction was used to confirm the geometry of trans-1,4-bis(4-amidinophenoxy)-2-butene dihydrochloride dihydrate (trans-butenamidine). trans-Butenamidine is a semirigid analogue of pentamidine that has demonstrated good anti-Pneumocystis carinii activity in rats. Molecular modeling studies revealed that unlike pentamidine or propamidine, trans-butenamidine does not discriminate between AT and TA sequences in its binding to the minor groove of DNA. Crystal data: [C18H22N4O2(2+)][Cl(-)]2[H2O]2, triclinic space group, P1, a = 9.443(1) A, b = 11.400(1) A, c = 11.919(1) A, alpha = 62.19(1) degree, beta = 81.10(1) degree, gamma = 72.19(1) degree, V = 1080.3(3) A3, Z = 2, R = 0.054 for 1149 observed reflections with I > 3 sigma (1).


Subject(s)
Pentamidine/analogs & derivatives , Pentamidine/chemistry , Base Sequence , Chemical Phenomena , Chemistry, Physical , Crystallography, X-Ray , Hydrogen Bonding , Models, Molecular , Molecular Sequence Data
8.
Biochemistry ; 33(34): 10271-85, 1994 Aug 30.
Article in English | MEDLINE | ID: mdl-8068666

ABSTRACT

Two-dimensional NMR methods were used to model the possible solution structure of an intercalative complex of 9-aminoellipticine (Aell), a polycyclic pyridocarbazolamine, covalently bound to an apurinic ring-opened deoxyribose site of a duplex DNA fragment in the reduced Schiff base form. The required oligonucleotide single strand containing covalently attached aminoellipticine was obtained by reductive amination in the presence of sodium cyanoborohydride. The combined NMR-energy minimization methods were employed to refine the model structures of two distinct forms, intrahelical and extrahelical, of a control 9-mer duplex DNA, d(CGTG.dr.GTGC).d(GCACTCACG), which contains an apurinic site positioned opposite a dT residue on the complementary strand. The model structure of an aminoellipticine conjugate with the same DNA sequence, derivatized via the aforementioned covalent attachment, was also obtained by incorporating intermolecular drug-DNA and intra- and internucleotide NOE-derived proton-proton distance estimates as restraints in energy minimization routines. The indole ring system of aminoellipticine, which is inserted at the apurinic site, intercalates between and is parallel to flanking GC base pairs. The pyridinic ring of aminoellipticine, in protonated form, also stacks between cytidine and thymidine bases on the complementary strand, which is consistent with the observation that the normal sequential NOE connectivity at the 5'-C13-T14 step is broken and indeed diverted through the ellipticine moiety, e.g., C13-Aell-T14 connectivities through the Aell-H4/C5Me protons. Interestingly, the partial stacking of the pyridinic ring is observed only between the 5'-CT step vs an adjacent 5'-TC step, owing to inherently weak stacking interactions associated with the former. In the absence of any potential groups that can participate in electrostatic or hydrogen-bonding interactions with the nucleic acid, pi-pi stacking and hydrophobic contacts at the intercalation site appear to be the important factors in determining stability and conformation of the aminoellipticine-DNA conjugate. Stacking interactions in such a bistranded intercalative complexation of aminoellipticine apparently govern the formation of a single intrahelical form of a right-handed B-type DNA duplex. The overall structural features lead us to propose working models for an enzyme-like DNA cleavage activity of 9-aminoellipticine and the observed inhibition of the AP endonuclease-dependent DNA excision-repair pathway.


Subject(s)
DNA/chemistry , Ellipticines/chemistry , Nucleic Acid Heteroduplexes/chemistry , Base Sequence , Binding Sites , DNA/chemical synthesis , DNA Repair/drug effects , Ellipticines/pharmacology , Intercalating Agents/chemistry , Magnetic Resonance Spectroscopy , Models, Molecular , Molecular Sequence Data , Molecular Structure , Nucleic Acid Heteroduplexes/chemical synthesis , Oligodeoxyribonucleotides/chemical synthesis , Oligodeoxyribonucleotides/chemistry , Schiff Bases/chemistry
9.
J Med Chem ; 35(16): 2995-3002, 1992 Aug 07.
Article in English | MEDLINE | ID: mdl-1501225

ABSTRACT

A template-directed approach to the design of a DNA-DNA interstrand cross-linker based upon the structure of a bis-tomaymycin-duplex adduct has been carried out. Tomaymycin is a member of the pyrrolo[1,4]benzodiazepines antitumor antibiotics. In a previous study (F.L. Boyd et al., Biochemistry 1990, 29, 2387-2403), we have shown that two tomaymycin molecules can be covalently bound to a 12-mer duplex molecule, where the drug molecules are on opposite strands six base-pairs apart, and the stereochemistry at the drug bonding site, and orientation in the minor groove, was defined by high-field NMR. This bis-tomaymycin 12-mer duplex adduct maintains the self-complementarity of the duplex and a B-type structure. In the present study we have shown using high-field NMR that this same 12-mer sequence can be truncated by two base pairs so that the two tomaymycin-modified guanines are now only four base-pairs apart, the two species of tomaymycin molecules are still bound with the same stereochemistry and orientation, and the 10-mer duplex adduct maintains its self-complementarity. In a second 10-mer duplex we have shown that changing the bonding sequence from 5'CGA to 5'AGC does not significantly affect the structure of the bis-tomaymycin-duplex adduct. However, when the sequence is rearranged so that the drugs point in a tail-to-tail orientation rather than in the previous head-to-head configuration, there are more than one species of tomaymycin bound to DNA, and, as a consequence, the bis-tomaymycin 10-mer duplex adduct loses its self-complementarity. Last, we have used the 10-mer duplex containing the 5'CGA sequence, in which the tomaymycin molecules are oriented head to head, to design an interstrand cross-linking species in which the two drug molecules are linked together with a flexible linker molecule.


Subject(s)
Cross-Linking Reagents/pharmacology , DNA/drug effects , Base Sequence , Benzodiazepinones/chemistry , Benzodiazepinones/pharmacology , Computer Simulation , Cross-Linking Reagents/chemistry , DNA/chemistry , Drug Design , Guanine/chemistry , Magnetic Resonance Spectroscopy , Models, Molecular , Molecular Sequence Data , Templates, Genetic
10.
Chem Res Toxicol ; 5(2): 167-82, 1992.
Article in English | MEDLINE | ID: mdl-1322736

ABSTRACT

The solution structure of the GC-rich non-self-complementary DNA 12-mer duplex (I), which contains a (+)-CC-1065 highly reactive bonding sequence 5'AGTTA* (where * denotes the [formula: see text] covalent modification site), has been examined thoroughly by one- and two-dimensional proton and phosphorus NMR spectroscopy, hydroxyl-radical footprinting, and NOESY restrained molecular mechanics and dynamics calculations. The assignments of the nonexchangeable proton resonances (except some of the H5' and H5" protons due to severe resonance overlap), phosphorus resonances, and the exchangeable resonances (except amino protons of adenosine and guanosine) of this 12-mer duplex have been made. The results show that this 12-mer duplex maintains an overall B-form DNA with all anti base orientation throughout in aqueous solution at room temperature. Hydroxyl-radical footprinting experiments on a 21-mer sequence that contains this 12-mer duplex used for NMR studies showed that the minor groove is somewhat narrowed at the 7G-8T and 17A-18C steps, as indicated by the inhibition of cleavage at these locations. Although both high-field NMR and hydroxyl-radical footprinting experiments supported a bent-like structure for this 12-mer duplex, nondenaturing gel electrophoresis on the ligated 21-mer sequence that contains this 12-mer duplex did not show the abnormally slow migration characteristic of a bent DNA duplex. Analysis of the NMR data sets reveals several local structural perturbations similar to those found on an (A)n tract DNA duplex. For example, the existence of a propeller twist was detected within the A.T-rich region for both the 12-mer and the (A)n tract DNA duplexes. The 18CH5 aromatic resonance that is directly adjacent to the 3' side of the 5'TAA segment was significantly shifted upfield with a chemical shift of 5.10 ppm, which is almost within the region normally associated with sugar H3' protons. The sugar geometries for 18C and 7G, which are located to the 3' side of the 5'TAA segment, are proposed to be in the neighborhood of C3'-endo and O1'-endo in equilibrium C3'-endo, respectively. We propose that this unusually upfield-shifted resonance signal for 18CH5 and the average C3'-endo sugar geometry for 18C nucleotide on the 12-mer duplex is connected with the peculiar conformation, possibly a transient kink, within the 5'AC/GT step. The results of the NOESY restrained molecular mechanics and dynamics calculations on the 12-mer sequence reveal two kinks, which are located on either side of the 18C nucleotide that has an average C3'-endo sugar geometry.(ABSTRACT TRUNCATED AT 400 WORDS)


Subject(s)
DNA/chemistry , Oligodeoxyribonucleotides/chemistry , Base Sequence , Hydroxides , Hydroxyl Radical , Magnetic Resonance Spectroscopy , Models, Molecular , Molecular Sequence Data , Protons
11.
J Med Chem ; 34(7): 1990-8, 1991 Jul.
Article in English | MEDLINE | ID: mdl-2066972

ABSTRACT

The binding of Saframycin A to the octanucleotide duplex d(GATGCATC)2 was investigated using molecular dynamics. For covalent binding at N2 of the central guanine, only the R configuration at the alkylating carbon (C7) was permitted for B DNA and the 3' direction in the minor groove was preferred by 50.6 kcal/mol. The dihydroquinone form of saframycin A gave stronger binding than the quinone, in agreement with the literature. Addition of solvent and counterions made no significant change in the geometry model. The proposed mechanism of DNA alkylation, involving iminium ion intermediates from the dihydroquinone or quinone, was investigated by modeling these species. They gave models with good net binding enthalpies, and C7 was in close proximity to N2 of guanine. The noncovalent binding of saframycin A and its dihydroquinone in the vicinity of guanine also was favorable in the 3' direction.


Subject(s)
Antibiotics, Antineoplastic/metabolism , Computer Simulation , Isoquinolines/metabolism , Models, Molecular , Oligonucleotides/metabolism , Polyribonucleotides/metabolism , Base Sequence , Molecular Conformation , Molecular Sequence Data , Stereoisomerism , Structure-Activity Relationship
12.
J Med Chem ; 34(7): 2079-88, 1991 Jul.
Article in English | MEDLINE | ID: mdl-2066979

ABSTRACT

Cyanocycline A was found to have a pKa of 6.6. Protonation of N14 was established by 1H NMR spectroscopy. In strongly acidic solution the oxazolidine ring opened irreversibly. A model was derived for the binding of naphthyridinomycin and cyanocycline A to the hexanucleotide duplex d(ATGCAT)2, by using the molecular mechanics and dynamics modules of AMBER 3.0. It involved protonation on the oxazolidine-ring nitrogen, reduction of the quinone ring to a hydroquinone, formation of an iminium ion with loss of the C7 substituent, noncovalent binding in the minor groove with the hydroquinone ring in the 3'-direction from guanine, and covalent binding to the 2-amino group of this guanine with C7 adopting the R configuration. This model is consistent with the experimental evidence on the DNA binding of these drugs. An alternative binding mode based on opening of the oxazolidine ring and alkylation at C3a also was feasible according to molecular mechanics calculations. The geometry of naphthyridinomycin does not permit interstrand cross-linking involving both C3a and C7, but formation of a cross-link to protein appears possible. When the covalent naphthyridinomycin-d(ATGCAT)2 models were refined in the presence of water and counterions, the models with the most favorable net binding enthalpies were the same as those produced by simulation in vacuum. Qualitative estimates of the relative entropy changes resulting from adduct formation were based on the number of ordered (hydrogen bonded) water molecules released from d(ATGCAT)2 and from the drug. In all cases but one, d(ATGCAT)2 loses five water molecules. It loses six in the C3a covalent model with 5',S geometry. Naphthyridinomycin hydroquinone loses up to two water molecules, depending on the particular adduct. The 3',R model was again favored for the C7 covalent adduct. Among the C3a covalent models, the one with 5',R geometry lost the second most water molecules, but it had the best binding enthalpy.


Subject(s)
Anti-Bacterial Agents/metabolism , Computer Simulation , DNA/metabolism , Models, Molecular , Oligoribonucleotides/metabolism , Alkylation , Base Sequence , Binding, Competitive , Chemical Phenomena , Chemistry , Magnetic Resonance Spectroscopy , Molecular Sequence Data , Naphthyridines/metabolism , Stereoisomerism , Structure-Activity Relationship
13.
Mol Biochem Parasitol ; 47(1): 119-28, 1991 Jul.
Article in English | MEDLINE | ID: mdl-1857381

ABSTRACT

We have isolated a cDNA clone corresponding to a single-copy nuclear gene that is upregulated at the mRNA level during in vitro differentiation of bloodstream trypomastigotes of strains of both Trypanosoma brucei brucei and Trypanosoma brucei rhodesiense to procyclic forms. Transcript levels begin to increase within minutes of introduction of bloodstream forms into culture and peak well before cultures exhibit a procyclic morphology. This increase in transcript levels was found to occur both in the absence of protein synthesis and in a nontransforming strain blocked very early in the developmental program, both conditions under which accumulation of procyclic acidic repetitive protein (PARP) transcripts did not occur in control experiments. DNA sequence analysis reveals an open reading frame sufficient to encode a protein of approximately 50 kDa within the cDNA, but data base searches for homology at either the amino acid or nucleotide level revealed no related sequences. A high density of kinase consensus target sites in the deduced amino acid sequence suggests that the gene product may be a phosphoprotein.


Subject(s)
Membrane Glycoproteins , Phosphoproteins/genetics , Trypanosoma brucei brucei/genetics , Variant Surface Glycoproteins, Trypanosoma/genetics , Amino Acid Sequence , Animals , Base Sequence , Cell Differentiation/genetics , DNA, Protozoan/chemistry , Genes , Kinetics , Molecular Sequence Data , Multigene Family , Phosphoproteins/biosynthesis , Protozoan Proteins/biosynthesis , Protozoan Proteins/genetics , RNA, Messenger/metabolism , Rats , Transcription, Genetic , Trypanosoma brucei brucei/growth & development , Up-Regulation , Variant Surface Glycoproteins, Trypanosoma/biosynthesis , Variant Surface Glycoproteins, Trypanosoma/isolation & purification
14.
J Med Chem ; 33(6): 1549-53, 1990 Jun.
Article in English | MEDLINE | ID: mdl-2342051

ABSTRACT

The relative DNA binding strengths of bisantrene and nine new analogues were measured by spectrophotometric titration and melt transition temperature (Tm) techniques. Data from the spectrophotometric titrations could not be fit by simple Scatchard plots. However, they were fit by a McGhee-von Hippel equation over part of the binding range. The entire range of data was fit by a smoothing cubic spline function. The first derivative of this function gave, for each compound, a curve whose intercept provided a measure of relative binding strength. The delta Tm values agreed qualitatively with the spectrophotometric titration results, although there was not a precise linear relationship. Determinations of macroscopic pKas revealed that most of the compounds were dications at pH 7.0, but a few were mixtures of monocations and dications. No correlation was found between these binding studies and antitumor potencies in a clonogenic assay, which suggests that factors other than DNA binding can determine cytotoxicity for some of the analogues.


Subject(s)
Antibiotics, Antineoplastic/metabolism , DNA/metabolism , Animals , Anthracenes/metabolism , Cattle , Drug Screening Assays, Antitumor , Structure-Activity Relationship
15.
J Parasitol ; 76(3): 445-7, 1990 Jun.
Article in English | MEDLINE | ID: mdl-2161918

ABSTRACT

Apocytochrome c was isolated from procyclic trypomastigotes of Trypanosoma rhodesiense EATRO 1895 and purified on Amberlite IRC-50 ion-exchange resin. Tryptic peptides were generated from the purified apoprotein and a partial amino acid sequence was determined. A comparison of the amino acid sequence of Crithidia fasciculata with the partial amino acid sequence of T. rhodesiense reveals significant homology.


Subject(s)
Crithidia/analysis , Cytochrome c Group/analysis , Trypanosoma brucei brucei/analysis , Amino Acid Sequence , Animals , Molecular Sequence Data
17.
J Comput Aided Mol Des ; 2(2): 91-106, 1988 Jul.
Article in English | MEDLINE | ID: mdl-3199155

ABSTRACT

Computer-based models were derived for the covalent and noncovalent binding of the antitumor antibiotic quinocarcin to a representative DNA segment, d(ATGCAT)2. They showed that a mode of action, involving opening of the oxazolidine ring to give an iminium ion, followed by initial noncovalent binding in the minor groove and subsequent alkylation of the 2-amino group of guanine, was rational and attended by favorable interaction energies in each step. The best model had the aryl ring of quinocarcin lying in the 3' direction from the covalent binding site and an R configuration at the carbon involved in covalent bond formation. It also showed that the preferred absolute configuration for quinocarcin was the reverse of that arbitrarily assigned in the literature.


Subject(s)
DNA/metabolism , Computer Simulation , DNA Damage , Isoquinolines/isolation & purification , Isoquinolines/metabolism , Models, Molecular , Nucleic Acid Conformation , Stereoisomerism
18.
Exp Parasitol ; 62(1): 85-91, 1986 Aug.
Article in English | MEDLINE | ID: mdl-3013670

ABSTRACT

One- and two-dimensional gel electrophoresis of the solubilized mitochondrial proteins of bloodstream and procyclic trypomastigote Trypanosoma brucei rhodesiense and radiolabeling of proteins in the presence of cycloheximide were used to identify proteins synthesized in the trypanosome mitochondrion. The proteins which comprise the mitochondrion were found to be very similar in both bloodstream and procyclic trypomastigotes, but do differ in their level of synthesis. A protein putatively identified as subunit II of cytochrome oxidase (EC 1.9.3.1) was detected in mitochondria from both the procyclic and bloodstream organisms. The presence of this protein in bloodstream trypomastigotes and the overall similarity of protein content in the trypanosome mitochondria is noteworthy in view of the fact that bloodstream trypomastigotes have a repressed mitochondrion with no detectable tricarboxylic acid cycle or cytochrome electron transport chain.


Subject(s)
Mitochondria/metabolism , Trypanosoma brucei brucei/metabolism , Animals , Antibodies, Monoclonal , Cycloheximide/pharmacology , Electron Transport Complex IV/metabolism , Electrophoresis, Polyacrylamide Gel , Mitochondria/drug effects , Molecular Weight , Proteins/isolation & purification , Proteins/metabolism , Rats , Rats, Inbred Strains , Trypanosoma brucei brucei/physiology
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