Autolysis at the disintegrin domain of patagonfibrase,a metalloproteinase from Philodryas patagoniensis (Patagonia Green Racer; Dipsadidae) venom

Patagonfibrase is a 57.5-kDa hemorrhagic metalloproteinase isolated from the venom of Philodryas patagoniensis (Patagonia Green Racer), a South American rear-fanged snake. Herein we demonstrate that patagonfibrase undergoes autolysis at its pH optimum (7.5) and at 37 °C, primarily producing a ∼ 32.6 kDa fragment composed of disintegrin-like and cysteine-rich domains, as identified by mass spectrometry and N-terminal sequencing. The autolysis site for production of this fragment is similar to that observed for metalloproteinases from front-fanged Viperidae snake venoms. In the presence of Ca²⁺, patagonfibrase was only partially autolysed, giving rise mainly to one fragment of ∼ 52.2 kDa. In addition, calcium markedly enhanced the azocaseinolytic activity of patagonfibrase. Our findings contribute to the understanding of the structural and mechanistic bases of this family of metalloenzymes that are widely distributed among snake venoms, demonstrating that important post-translational modifications such as proteolysis can also contribute to the diversity and complexity of proteins found in rear-fanged snake venoms.     

Richness,geographic distribution patterns,and areas of endemism of selected angiosperm groups in Mexico

Mexico is a megadiverse country. Presently, 22 126 species of angiosperms have been registered within its territory and 11 001 are considered to be endemic. However, their geographical distributions are far from homogeneous. In addition, Mexico is the center of diversification of several groups. Our analysis focused on such groups. The aims were to identify areas of species richness and endemism. A data matrix with 766 species and 25 579 geographical records was analyzed. It included Calochortus (Liliaceae); Bletia (Orchidaceae); Tigridieae (Iridaceae); Amaryllidaceae; Poliantheae, Echeandia (Asparagaceae); Crassulaceae; Hylocereus (Cactaceae); Solanum, Lycianthes and Physalinae (Solanaceae); Salvia section Membranaceae (Lamiaceae); and Cosmos and Dahlia (Asteraceae). Using Geographic Information Systems, we determined richness and distribution based on: (i) Mexican political divisions, (ii) biogeographical regions and provinces, (iii) a grid of 0.5 × 0.5° cells, and (iv) elevation. The areas of endemism were estimated using the endemicity analysis. The highest number of taxa and endemic plants were concentrated within the Transmexican Volcanic Belt in the Mexican Transition Zone. This mountain range has been recognized as a province on the basis of geologic, tectonic, geomorphologic, physiographic and biogeographic criteria. It is a 1000 km long volcanic arc that extends east to west through Central Mexico and is variably from 80 to 230 km wide, between 17°30′ to 20°25′N and 96°20′ to 105°20′W. Our results represent a local deviation from the global richness latitudinal gradient of angiosperm species.     

Two new species of Physalis (Solanaceae) endemic to Jalisco, Mexico

Two new species ofPhysalis, both endemic to Jalisco, Mexico, are described and illustrated:P. longiloba andP. tamayoi.

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Resumen  Se describen e ilustran dos nuevas especies dePhysalis endémicas de Jalisco, México:P. longiloba y.P. tamayoi.

     

Photoreactivation of superoxide dismutase by intensive red (laser) light

The effects of helium-neon laser (HNL) on activity, absorption spectra, and ESR signals of superoxide dismutase (SOD; E Cul. 15.1.1) from bovine erythrocytes in acid medium were investigated. It was found that incubation during 2 hours at pH 5.9 led to inactivation of the enzyme. The subsequent illumination of SOD by HNL brought about the enzyme reactivation. Both absorption and ESR spectra were changed after incubation at pH 5.9 and restored after laser irradiation. In a model system, copper-histidine complex, absorption maximum was shifted from 632–633 nm at pH 5.8 to 639–640 nm at pH 8.5–9.0. The similar shift of the maximum was observed after illumination by HNL at pH 5.8. It may be postulated that the photoreactivation of SOD consists essentially in deprotonation of His-61 residue in the enzyme active site and subsequent recovery of imidasol bridge between copper and zinc which had been destroyed at low pH.

Since many other enzymes possess similar histidine-copper structures in their active sites, one may expect diverse effects of red (laser) light on the enzyme activity. Heme-containing enzyme, catalase was also found to be photoreactivated by HNL after inactivation at pH 6.0.     

Simultaneous determination of Fe(III) and Fe(II) in water solutions and tissue homogenates using desferal and 1,10-phenanthroline

At neutral pH values 1,10-phenanthroline forms a colored complex with Fe(II), but it does not form such a complex with Fe(III). On the contrary, only Fe(III) forms with desferal a yellow complex with a g = 4.3 electron paramagnetic resonance (EPR) signal, but Fe(II) is rapidly oxidized by desferal to Fe(III), which gives then a yellow complex. On the basis of these facts, a method for simultaneous determination of both Fe(II) and Fe(III) ions was elaborated using a desferal-phenanthroline mixture. Two ways of detecting Fe(II) and Fe(III) have been suggested: (1) the spectrophotometric method for transparent water solutions, and (2) the EPR-spectrometric method for turbid solutions and tissue homogenates. The latter method was applied for determination of free and weakly bound iron in rat liver. The Fe(II) amount in intact liver was 22.2 ± 7.6 nmol/g of wet tissue; free Fe(III) was not found.     

Use of NMR metabolomics to analyze the targets of D-cycloserine in mycobacteria: role of D-alanine racemase

D-Cycloserine (DCS) is only used with multidrug-resistant strains of tuberculosis because of serious side effects. DCS is known to inhibit cell wall biosynthesis, but the in vivo lethal target is still unknown. We have applied NMR-based metabolomics combined with principal component analysis to monitor the in vivo effect of DCS on Mycobacterium smegmatis. Our analysis suggests DCS functions by inhibiting multiple protein targets.     

Genetic diversity within and among two Argentinean and one Mexican species of Acacia (Fabaceae)

Acacia is a pantropical genus comprising > 1450 species. Following Vassal's treatment Acacia is considered as a single genus with three subgenera (Acacia, Aculeiferum and Phyllodineae). Acacia caven, A. curvifructa and A. farnesiana belong to subgenus Acacia and the relationship between them is controversial. The aim of this study was to elucidate the relationship between the three species using amplified fragment length polymorphism, analysing 15 populations of these species, and to compare the results obtained with those from a morphological analysis. Genetic diversity indices (percentage of polymorphic loci, genetic diversity) showed that genetic variation in A. caven is higher than that in A. curvifructa and A. farnesiana. Of the total genetic diversity in A. caven and A. farnesiana, most is found within populations (∼70%). Analysis with STRUCTURE showed that the optimal number of clusters (K) was ten, and in all cases where populations were grouped they were geographically close and/or belong to the same variety. The morphological canonical discriminant analysis did not result in a separation between all individuals, indicating that they do not harbour consistent morphological discontinuities. Altogether, the results of our molecular analyses showed the existence of significant differences between A. caven, A. curvifructa and A. farnesiana, which argues for recognizing them as different species. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2015, 177 , 593–606.     

Do assortative mating and immigrant inviability help maintain population genetic structuring of an herbivore on a crop and a wild relative?

Population genetic structuring is common among herbivorous insects and frequently is associated with divergent host plants, such as crops and their wild relatives. Previous studies showed population genetic structuring in corn leafhopper Dulbulus maidis in Mexico, such that the species consists of two sympatric, host plant-associated populations: an abundant and widespread "pestiferous” population on maize (Zea mays mays), and a small and localized "wild" population on perennial teosinte (Zea diploperennis). a maize wild relative with a limited distribution. This study addressed whether assortative mating and immigrant inviability mediate genetic structuring of corn leafliopper by comparing the mating and reproductive successes of pestiferous and wild females that colonize their nonassociated host plants against the successes of females colonizing their associated host plants. Assortative mating was assessed by comparing mating frequencies and premating and mating times among females of each population on each host plant: immigrant inviability was assessed by comparing, across two generations, the fecundity, survival, development time, sex ratio, and population growth rate among leafhopper populations and host plants. Our results showed that on maize, and compared to resident, pestiferous females, wild females were more likely to mate, and greater proportions of their offspring survived to adult stage and were daughters;consequently, the per-generation population growth rate on maize was greater for immigrant, wild leafhoppers compared to resident, pestiferous leafhoppers. Our results suggested that wild leafhoppers emigrating to maize have a fitness advantage over resident, pestiferous leafhoppers, while immigrant pestiferous and resident wild leafhoppers on teosinte have similar fitnesses.