Glycoproteins of trypanosomes: their biosynthesis and biological significance

1. Trypanosomes are unicellular parasites that cause human sleeping sickness in Africa and Chagas' disease in South America. Glycoproteins are important components of their plasma membrane. 2. The bloodstream form of the extracellular salivarian African trypanosome (e.g. Trypanosoma brucei) has the ability to express on its cell surface a repertoire of variant surface glycoproteins (VSGs) and in so doing, evades the immune response of the host (antigenic variation). 3. The VSG is probably synthesized initially in a manner like that of the membrane-bound glycoproteins of mammalian systems, but it also undergoes some novel post-translational modifications. 4. The stercorarian South American trypanosome (Trypanosoma cruzi) is an intracellular parasite which expresses different glycoproteins on its plasma membrane at various stages of its life-cycle, but does not exhibit antigenic variation. 5. The biosynthesis and functions of trypanosomal glycoproteins are compared with those of mammalian glycoproteins, and are discussed with particular reference to potential targets for chemotherapy and immunotherapy of trypanosomiasis.     

The Redox Biochemistry of Protein Sulfenylation and Sulfinylation

Controlled generation of reactive oxygen species orchestrates numerous physiological signaling events (Finkel, T. (2011) Signal transduction by reactive oxygen species. J. Cell Biol. 194, 7–15). A major cellular target of reactive oxygen species is the thiol side chain (RSH) of Cys, which may assume a wide range of oxidation states (i.e. −2 to +4). Within this context, Cys sulfenic (Cys-SOH) and sulfinic (Cys-SO₂H) acids have emerged as important mechanisms for regulation of protein function. Although this area has been under investigation for over a decade, the scope and biological role of sulfenic/sulfinic acid modifications have been recently expanded with the introduction of new tools for monitoring cysteine oxidation in vitro and directly in cells. This minireview discusses selected recent examples of protein sulfenylation and sulfinylation from the literature, highlighting the role of these post-translational modifications in cell signaling.     

Turnover of Brain Monoamine Oxidase Measured In Vivo by Positron Emission Tomography Using l-[11C]Deprenyl

The distribution of carbon-11-labeled L-deprenyl, an irreversible inhibitor of monoamine oxidase type B (MAO-B), was determined in the baboon brain by positron emission tomography. The irreversible blood-to-brain transfer constant (influx constant, K_i) was measured using a complete metabolite-corrected arterial plasma concentration curve. This influx constant was used as a measure of functional enzyme activity for sequential determinations of MAO-B recovery following a single high dose of unlabeled l -deprenyl. The half-life for turnover of MAO-B was thus determined to be 30 days. Using appropriate irreversible inhibitors, this procedure should be generally useful for determining enzyme turnover rates in any organ in vivo and can be applied to some human studies as well.     

DNA polymorphism of human HLA-linked complement C4 allotypes, including C4 null alleles, in the Finnish population

Human HLA-linked complement C4 gene products, C4A and C4B, show extensive genetic polymorphism. In both loci, an allele without a gene product, C4 null, is also observed. We have performed a restriction enzyme analysis of genomic DNA samples from individuals having all common (frequency over 1%) C4 protein allotypes observed in the Finnish population. Only one allotype-specific RFLP marker was observed. With some enzymes a DNA polymorphism was observed, which was not detectable by C4 protein typing. Analysis of 10 different C4B null haplotypes and 4 C4A null haplotypes suggested that only one haplotype, HLA-B8 C4A0 B1, carried a C4A gene deletion. This was observed in all 4 unrelated individuals homozygous for this haplotype.     

An automated assay for adenylate cyclase using reversed-phase high-performance liquid chromatography

An automated high-performance liquid chromatographic method for the assay of 3',5'-cyclic AMP was developed using octylsilica. Total analysis time was 10.1 min, with cAMP eluting at 3 min. As little as 10 pmol of cyclic AMP could be detected by absorption at 260 nm. Peak height and area were linearly related to cyclic AMP concentration over at least two orders of magnitude. The analytical procedure gave good results in the assay of crude microsomal preparations of adenylate cyclase from both bovine brain and sea urchin eggs. The method was used to demonstrate that sea urchin adenylate cyclase is a Ca2+-activated enzyme.     

Cloning and sequencing of the peroxisomal amine oxidase gene from Hansenula polymorpha

We have cloned the AMO gene, encoding the microbody matrix enzyme amine oxidase (EC 1.4.3.6) from the yeast Hansenula polymorpha. The gene was isolated by differential screening of a cDNA library, immunoselection, and subsequent screening of a H. polymorpha genomic library. The nucleotide sequence of a 3.6 kilobase stretch of DNA containing the amine oxidase (AMO) gene was determined. The AMO gene contains an open reading frame of 692 amino acids, with a relative molecular mass of 77,435. The 5' and 3' ends of the gene were mapped and show that the transcribed region measures 2134 nucleotides. The derived amino-acid sequence was confirmed by sequencing an internal proteolytic fragment of the purified protein. Amine oxidase contains the tripeptide sequence Ser-Arg-Leu, located 9 residues from the carboxy terminus, which may represent the topogenic signal for protein import into microbodies.     

An extract of Commiphora erythraea: a repellent and toxicant against ticks

A hexane extract of the gum of an African plant, Commiphora erythraea Engler (Burseraceae), has larvicidal and repellent activity against the lone star tick, Amblyomma americanum (L.) and the American dog tick, Dermacentor variabilis (Say). Adult deer ticks, Ixodes dammini Spielman, Clifford, Piesman and Corwin, were also repelled by the extract. Concentrations of 0.02 mg/cm² of the extract impregnated onto filter paper killed 96.15 (±3.56)% of A. americanum larvae exposed to it for 24h. A concentration of 0.16 mg/cm² was needed to kill 80.3% of D. variabilis larvae. The extract was less effective as a larvicide against A. americanum and D. variabilis than permethrin. Less than 15.5% of A. americanum larvae and adults and D. variabilis and I. dammini adults entered and remained for 2 or 3 min on areas of cloth strips treated with the extract at the rate of 0.2 mg/cm². However, 73.3 to 83.3% of the ticks tested entered and remained in areas treated with hexane. Permethrin was about 1 or 2 orders of magnitude more effective against A. americanum larvae as a repellent than the extract.

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Résumé Un extrait à l'hexane de la gomme de la plante africaine, C. erythraea Engler (Burseraceae) a des effets larvicides et répulsifs contre les tiques, Amblyomma americanum L. et Dermacentor variabilis Say; les adultes de Ixodes dammini Spielman, Clifford, Piesman & Corwin, ont été aussi éloignés par l'extrait. Une concentration de 0,02 mg/cm² de l'extrait imprégnant du papier filtre a tué 96,15% (±3,56) larves de A. americanum exposées pendant 24 h. Une concentration de 0,16 mg/cm² a été nécessaire pour tuer 80,3% des larves de D. variabilis. L'extrait a été moins efficace que la perméthrine comme larvicide contre A. americanum et D. variabilis. Moins de 15,5% des larves et des adultes de A. americanum et des adultes de D. variabilis et de I. dammini pénétrèrent et séjournèrent 2 à 3 minutes dans des morceaux de tissu traités avec l'extrait à raison de 0,2 mg/cm², contre 73,3 et 83,3% qui pénétrèrent et restèrent lors de traitement avec de l'hexane. Contre A. americanum, la perméthrine est un répulsif d'un ordre de grandeur une à deux fois supérieur à l'extrait de C. erythraea.

     

Biochemical Studies of Paraquat-Tolerant Mutants of the Fern Ceratopteris richardii

Enzymes and metabolites associated with mitigation of paraquat toxicity were compared in two paraquat-tolerant mutants and a sensitive wild-type strain of the fern Ceratopteris richardii Brongn. In 21-day-old gametophytes, the specific activities of superoxide dismutase, catalase, peroxidase, glutathione reductase, dehydroascorbate reductase, and ascorbate peroxidase showed no differences that would explain mutant tolerance. Constitutive levels of ascorbate and glutathione also did not differ significantly in the three strains. An experiment testing the inducibility of paraquat tolerance revealed no change in the dose response of mutant or wild type gametophytes after exposure to sublethal concentrations of the herbicide. Uptake of paraquat by whole gametophytes was also equivalent in mutants and wild type. These data suggest that the physiological basis for tolerance in these mutants, unlike several other tolerant biotypes reported, does not lie in the oxygen radical scavenging system, in an inducible stress response, or in a block to whole-plant uptake.