RESUMO
The relationship between protein stability and functional evolution is little explored in proteins purified from natural sources. Here, we investigated a novel family of egg proteins (Perivitellin-1, PV1) from Pomacea snails. Their remarkable stability and clade-related functions in most derived clades (Canaliculata and Bridgesii) make them excellent candidates for exploring this issue. To that aim, we studied PV1 (PpaPV1) from the most basal lineage, Flagellata. PpaPV1 displays unparalleled structural and kinetic stability, surpassing PV1s from derived clades, ranking among the most hyperstable proteins documented in nature. Its spectral features contribute to a pale egg coloration, exhibiting a milder glycan binding lectin activity with a narrower specificity than PV1s from the closely related Bridgesii clade. These findings provide evidence for substantial structural and functional changes throughout the genus' PV1 evolution. We observed that structural and kinetic stability decreased in a clade-related fashion and was associated with large variations in defensive traits. For instance, pale PpaPV1 lectin turns potent in the Bridgesii clade, adversely affecting gut morphology, while giving rise to brightly colored PV1s providing eggs with a conspicuous, probably warning signal in the Canaliculata clade. This work provides a comprehensive comparative analysis of PV1s from various apple snail species within a phylogenetic framework, offering insights into the interplay among their structural features, stability profiles and functional roles. More broadly, our work provides one of the first examples from natural evolution showing the crucial link among protein structure, stability and evolution of new functions.
Assuntos
Proteínas do Ovo , Filogenia , Caramujos , Animais , Caramujos/genética , Caramujos/fisiologia , Caramujos/química , Proteínas do Ovo/genética , Proteínas do Ovo/química , Proteínas do Ovo/metabolismo , Estabilidade Proteica , Óvulo/química , Óvulo/metabolismoRESUMO
The absence of robust interspecific isolation barriers among pantherines, including the iconic South American jaguar (Panthera onca), led us to study molecular evolution of typically rapidly evolving reproductive proteins within this subfamily and related groups. In this study, we delved into the evolutionary forces acting on the zona pellucida (ZP) gamete interaction protein family and the sperm-oocyte fusion protein pair IZUMO1-JUNO across the Carnivora order, distinguishing between Caniformia and Feliformia suborders and anticipating few significant diversifying changes in the Pantherinae subfamily. A chromosome-resolved jaguar genome assembly facilitated coding sequences, enabling the reconstruction of protein evolutionary histories. Examining sequence variability across more than 30 Carnivora species revealed that Feliformia exhibited significantly lower diversity compared to its sister taxa, Caniformia. Molecular evolution analyses of ZP2 and ZP3, subunits directly involved in sperm-recognition, unveiled diversifying positive selection in Feliformia, Caniformia and Pantherinae, although no significant changes were linked to sperm binding. Structural cross-linking ZP subunits, ZP4 and ZP1 exhibited lower levels or complete absence of positive selection. Notably, the fusion protein IZUMO1 displayed prominent positive selection signatures and sites in basal lineages of both Caniformia and Feliformia, extending along the Caniformia subtree but absent in Pantherinae. Conversely, JUNO did not exhibit any positive selection signatures across tested lineages and clades. Eight Caniformia-specific positive selected sites in IZUMO1 were detected within two JUNO-interaction clusters. Our findings provide for the first time insights into the evolutionary trajectories of ZP proteins and the IZUMO1-JUNO gamete interaction pair within the Carnivora order.
Assuntos
Caniformia , Carnívoros , Panthera , Animais , Masculino , Receptores de Superfície Celular/genética , Proteínas do Ovo/genética , Proteínas do Ovo/química , Proteínas do Ovo/metabolismo , Sêmen/metabolismo , Interações Espermatozoide-Óvulo/genética , Carnívoros/genética , Caniformia/metabolismo , Feliformes/metabolismo , Panthera/metabolismo , Zona Pelúcida/metabolismoRESUMO
This study aimed to examine the impact of levels of dietary supplementation with fat-soluble vitamins on the production performance and egg quality of laying hens. Three hundred Hy-Line White W-36 laying hens were evaluated from 28 to 44 weeks of age. The birds were allotted to one of six treatments in a randomized block design with 10 replicates with five birds each. Performance and egg quality parameters were evaluated in four 28-day periods. A corn and soybean meal-based basal diet was formulated so as to meet the nutritional requirements of the animals, with the exception of fat-soluble vitamins. The treatments consisted of dietary supplementation with 0%, 33.3%, 66.7%, 100.0%, 133.3% or 166.7% of fat-soluble vitamins (100% supplementation consisted of 7500 IU, 2000 IU, 10 IU and 1.8 mg of vitamins A, D3, E and K per kilogram of diet, respectively). Eggshell weight, shell thickness, shell strength, feed intake, egg weight, feed conversion per egg mass and feed conversion per dozen eggs showed a quadratic response (p0.05) to the treatments, whereas egg mass responded linearly. Optimal results were obtained at an average fat-soluble vitamin supplementation level of 109%, which corresponds to 8175 IU of vitamin A, 2180 IU of vitamin D3, 10.9 IU of vitamin E and 1.96 mg of vitamin K per kilogram of diet.(AU)
Assuntos
Animais , Galinhas/crescimento & desenvolvimento , Galinhas/fisiologia , Proteínas do Ovo/análise , Proteínas do Ovo/química , Ovos/análise , Suplementos Nutricionais/análiseRESUMO
This study aimed to examine the impact of levels of dietary supplementation with fat-soluble vitamins on the production performance and egg quality of laying hens. Three hundred Hy-Line White W-36 laying hens were evaluated from 28 to 44 weeks of age. The birds were allotted to one of six treatments in a randomized block design with 10 replicates with five birds each. Performance and egg quality parameters were evaluated in four 28-day periods. A corn and soybean meal-based basal diet was formulated so as to meet the nutritional requirements of the animals, with the exception of fat-soluble vitamins. The treatments consisted of dietary supplementation with 0%, 33.3%, 66.7%, 100.0%, 133.3% or 166.7% of fat-soluble vitamins (100% supplementation consisted of 7500 IU, 2000 IU, 10 IU and 1.8 mg of vitamins A, D3, E and K per kilogram of diet, respectively). Eggshell weight, shell thickness, shell strength, feed intake, egg weight, feed conversion per egg mass and feed conversion per dozen eggs showed a quadratic response (p0.05) to the treatments, whereas egg mass responded linearly. Optimal results were obtained at an average fat-soluble vitamin supplementation level of 109%, which corresponds to 8175 IU of vitamin A, 2180 IU of vitamin D3, 10.9 IU of vitamin E and 1.96 mg of vitamin K per kilogram of diet.
Assuntos
Animais , Galinhas/crescimento & desenvolvimento , Galinhas/fisiologia , Ovos/análise , Proteínas do Ovo/análise , Proteínas do Ovo/química , Suplementos Nutricionais/análiseRESUMO
There are many diseases linked to oxidative stress, including cancer. Importantly, endogenous antioxidants are insufficient to protect against this process. Peptides derived from food proteins produced by hydrolysis have been investigated as exogenous antioxidants. The present study aimed to identify novel peptides with antioxidant potential produced from egg and milk proteins hydrolysis with two new fungal proteases isolated from Eupenicillium javanicum and Myceliophthora thermophila. The degree of hydrolysis at several time points was calculated and correlated to DPPH scavenging and metal chelating assays, all hydrolysates presented antioxidant activity. Casein hydrolyzed by the M. thermophila protease showed the best antioxidant activity. The identified sequences showed that the proportions of amino acids that influence antioxidant activity support the antioxidant assay. Our data reveal the conditions necessary for the successful generation of antioxidant peptides using two novel fungal proteases. This opens a potential new avenue for the design and manufacture of antioxidant molecules.
Assuntos
Albuminas/química , Antioxidantes/química , Caseínas/química , Proteínas do Ovo/química , Peptídeos/química , Proteínas do Soro do Leite/química , Albuminas/farmacologia , Animais , Antioxidantes/farmacologia , Caseínas/farmacologia , Proteínas do Ovo/farmacologia , Eupenicillium/enzimologia , Peptídeo Hidrolases/química , Peptídeos/farmacologia , Proteólise , Sordariales/enzimologia , Proteínas do Soro do Leite/farmacologiaRESUMO
Devido a uma carência de informações técnico-cientificas sobre o efeito da carga de gemas em relação a interceptação da radiação solar, e o seu efeito na fertilidade de gemas, tem-se como objetivo desse trabalho avaliar o efeito do aumento da carga de gemas da videira Cabernet Franc em relação a interceptação da radiação fotossinteticamente ativa e o seu efeito na fertilidade de gemas em região de elevada altitude de Santa Catarina. Este experimento foi conduzido durante safra 2016/2017, em um vinhedo comercial, localizado no munícipio de São Joaquim. Utilizaram-se plantas de Cabernet Franc enxertadas sobre o porta-enxerto Paulsen 1103. Os tratamentos consistiram em quatro diferentes níveis de cargas de gemas: 15, 30, 50 e 75 gemas planta-1 . O delineamento experimental utilizado foi o de blocos ao acaso, com quatro blocos e cinco plantas por parcela. Os dados foram submetidos à análise de variância (ANOVA) e comparados pelo Teste Tukey a 5% de probabilidade de erro. Os resultados desse estudo evidenciam a importância da interceptação da radiação fotossinteticamente ativa na fertilidade de gemas, observando-se redução da fertilidade de gemas em situações de redução da interceptação da radiação fotossinteticamente ativa, nesse caso, causado pelo aumento da carga de gemas, que resulta em um maior número de ramos, proporcionando um dossel mais denso, reduzindo a interceptação da radiação fotossinteticamente ativa que alcança as gemas da videira, sendo esse efeito, mais pronunciado nas gemas localizada na posição basal do ramo.
Due to a lack of technical-scientific information about the effect of the retained node number on the interception of the solar radiation of the nodes and its effect on the fertility of nodes, this work aims to evaluate the effect of the increased number of nodes of the grapevine Cabernet Franc in relation to the interception of the photosynthetically active composition and its effect on the node fertility in high altitude region of Santa Catarina State. This experiment was conducted during the 2016/2017 harvest in a commercial vineyard located in the city of São Joaquim. Cabernet Franc plants grafted on the 'Paulsen 1103' rootstock were used. The treatments consisted of four different levels of retained node numbers: 15, 30, 50, and 75 nodes per plant. The experimental design was a randomized block design with four blocks and five plants per plot. The data were submitted to analysis of variance (ANOVA) and compared by the Tukey test with a 5% probability of error. The results of this study evidenced the importance of the interception of the photosynthetically active radiation in the nodes fertility, observing the reduction of the nodes fertility in situations of reduction of the interception of the photosynthetically active radiation, in this case, caused by the increase of the number of nodes, which resulted in a higher shoot number and branches, resulting in a denser canopy, and consequently reducing the interception of the photosynthetically active radiation that reaches the grape nodes, being this effect more pronounced in the buds located at the basal position of the branch.
Assuntos
Proteínas do Ovo/efeitos adversos , Proteínas do Ovo/química , Vitis/crescimento & desenvolvimento , Vitis/efeitos dos fármacos , Fármacos para a Fertilidade/análise , Vinho/análiseRESUMO
Devido a uma carência de informações técnico-cientificas sobre o efeito da carga de gemas em relação a interceptação da radiação solar, e o seu efeito na fertilidade de gemas, tem-se como objetivo desse trabalho avaliar o efeito do aumento da carga de gemas da videira Cabernet Franc em relação a interceptação da radiação fotossinteticamente ativa e o seu efeito na fertilidade de gemas em região de elevada altitude de Santa Catarina. Este experimento foi conduzido durante safra 2016/2017, em um vinhedo comercial, localizado no munícipio de São Joaquim. Utilizaram-se plantas de Cabernet Franc enxertadas sobre o porta-enxerto Paulsen 1103. Os tratamentos consistiram em quatro diferentes níveis de cargas de gemas: 15, 30, 50 e 75 gemas planta-1 . O delineamento experimental utilizado foi o de blocos ao acaso, com quatro blocos e cinco plantas por parcela. Os dados foram submetidos à análise de variância (ANOVA) e comparados pelo Teste Tukey a 5% de probabilidade de erro. Os resultados desse estudo evidenciam a importância da interceptação da radiação fotossinteticamente ativa na fertilidade de gemas, observando-se redução da fertilidade de gemas em situações de redução da interceptação da radiação fotossinteticamente ativa, nesse caso, causado pelo aumento da carga de gemas, que resulta em um maior número de ramos, proporcionando um dossel mais denso, reduzindo a interceptação da radiação fotossinteticamente ativa que alcança as gemas da videira, sendo esse efeito, mais pronunciado nas gemas localizada na posição basal do ramo.(AU)
Due to a lack of technical-scientific information about the effect of the retained node number on the interception of the solar radiation of the nodes and its effect on the fertility of nodes, this work aims to evaluate the effect of the increased number of nodes of the grapevine Cabernet Franc in relation to the interception of the photosynthetically active composition and its effect on the node fertility in high altitude region of Santa Catarina State. This experiment was conducted during the 2016/2017 harvest in a commercial vineyard located in the city of São Joaquim. Cabernet Franc plants grafted on the 'Paulsen 1103' rootstock were used. The treatments consisted of four different levels of retained node numbers: 15, 30, 50, and 75 nodes per plant. The experimental design was a randomized block design with four blocks and five plants per plot. The data were submitted to analysis of variance (ANOVA) and compared by the Tukey test with a 5% probability of error. The results of this study evidenced the importance of the interception of the photosynthetically active radiation in the nodes fertility, observing the reduction of the nodes fertility in situations of reduction of the interception of the photosynthetically active radiation, in this case, caused by the increase of the number of nodes, which resulted in a higher shoot number and branches, resulting in a denser canopy, and consequently reducing the interception of the photosynthetically active radiation that reaches the grape nodes, being this effect more pronounced in the buds located at the basal position of the branch.(AU)
Assuntos
Proteínas do Ovo/efeitos adversos , Proteínas do Ovo/química , Vitis/efeitos dos fármacos , Vitis/crescimento & desenvolvimento , Fármacos para a Fertilidade/análise , Vinho/análiseRESUMO
The complexation between lysozyme/carrageenan and ovalbumin/carrageenan was studied in situ using acidification. The complexes were analyzed in solutions with different NaCl concentrations and different protein/polysaccharide ratios. As the protein/polysaccharide ratio increased from 1:1 to 10:1, critical structure forming events (i.e., those associated with soluble, insoluble and large insoluble complexes) shifted to higher pH values for ovalbumin/carrageenan followed by decrease of G' values at ratios of 5:1 and 10:1. The increase in the ratio of lysozyme/carrageenan complexes suppressed the critical pH transition points that led to the formation of large insoluble complexes from pH 12.0 until 1.0, and the values of G' increased simultaneously, reaching the highest value at a ratio of 10:1. Addition of salt to the ovalbumin/carrageenan and lysozyme/carrageenan mixtures suppressed the electrostatic interaction between proteins and carrageenan at lower pH values and the critical pH transitions points, whereas at a ratio of 3:1 with a 0.01â¯M concentration, the coacervate yield of the complex reached 79.6%⯱â¯0.6 and 93.7%⯱â¯4.8 for the ovalbumin and lysozyme complexes, respectively. The rheological data associated with microscopy images show that interpolymer complexes with heterogeneous structures were formed for both complexes, and we suggest that complexes have a great potential to improve or extend the texture, mechanical stability, consistency, and taste of food products.
Assuntos
Carragenina/química , Carragenina/metabolismo , Proteínas do Ovo/química , Proteínas do Ovo/metabolismo , Reologia , Relação Dose-Resposta a Droga , Concentração de Íons de Hidrogênio , Concentração Osmolar , Ovalbumina/metabolismo , Pectinas/metabolismo , Ligação Proteica/efeitos dos fármacos , Cloreto de Sódio/farmacologia , Solubilidade , TermodinâmicaRESUMO
In contrast with vitellogenin maturation, it is unknown whether gastropod perivitellin precursors are subject to large structural changes. The gastropod reproductive tract includes an accessory organ, the albumen gland (AG), that produces and secretes perivitelline fluid. In the apple snail Pomacea canaliculata, the large, reddish-pink AG provides eggs with perivitellins that are defensive against predators. Although the AG makes a considerable contribution to apple snail biomass, field observations indicate that it is rejected by avian and mammalian predators, although the underlying reason remains unknown. By analyzing the structure-function properties of P. canaliculata perivitellin precursors, we provide insight into perivitellin maturation and its relationship with apple snail predator feeding behavior. Structural analysis using small-angle X-ray scattering, absorption and fluorescence spectroscopy, circular dichroism, electrophoresis, chromatography, and partial proteolysis showed that the size, shape, and structure of perivitellin precursors resemble those of egg mature forms. Functional analysis indicates that the precursors of the defensive perivitellins ovorubin (PcOvo) and perivitellin-2 (PcPV2) are highly stable and antinutritive, withstanding proteinase digestion and displaying structural stability of their quaternary structure under a wide pH range (4.0-10.0). Furthermore, AG extracts limit a predator's ability to digest nutrients and are toxic to mice (median lethal concentration 96 h after administration: 5.9 mg/kg). Treated mice displayed neurologic signs similar to those produced by egg PcPV2. Results indicate that apple snails store active precursors of egg proteins inside the AG, providing evidence that gastropod perivitellin precursors do not experience the large structural processing of invertebrate vitellogenin maturation. These defensive proteins provide the apple snail AG with neurotoxic, antinutritive, and antidigestive activity, a likely explanation for the predators' feeding behavior.
Assuntos
Líquidos Corporais/química , Proteínas do Ovo/química , Proteínas do Ovo/metabolismo , Óvulo/química , Caramujos/fisiologia , Animais , Concentração de Íons de Hidrogênio , Comportamento PredatórioRESUMO
The mammalian oocyte is surrounded by a matrix called the zona pellucida (ZP). This envelope participates in processes such as acrosome reaction induction, sperm binding and may be involved in speciation. In cat (Felis catus), this matrix is composed of at least three glycoproteins called ZP2, ZP3, and ZP4. However, recent studies have pointed to the presence of a fourth protein in several mammals (rat, human, hamster or rabbit), meaning that a reevaluation of cat ZP is needed. For this reason, the objective of this research was to analyze the protein composition of cat ZP by means of proteomic analysis. Using ZP from ovaries and oocytes, several peptides corresponding to four proteins were detected, yielding a coverage of 33.17%, 71.50%, 50.23%, and 49.64% for ZP1, ZP2, ZP3, and ZP4, respectively. Moreover, the expression of four genes was confirmed by molecular analysis. Using total RNA isolated from cat ovaries, the complementary deoxyribonucleic acids encoding cat ZP were partially amplified by reverse-transcribed polymerase chain reaction. Furthermore, ZP1 was totally amplified for the first time in this species. As far as we are aware, this is the first study that confirms the presence of four proteins in cat ZP.