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Don Trinh Nguyen Jens Christian G?pfert Nobuhiro Ikezawa Gillian MacNevin Meena Kathiresan Jürgen Conrad Otmar Spring Dae-Kyun Ro 《The Journal of biological chemistry》2010,285(22):16588-16598
Sesquiterpene lactones are characteristic natural products in Asteraceae, which constitutes ∼8% of all plant species. Despite their physiological and pharmaceutical importance, the biochemistry and evolution of sesquiterpene lactones remain unexplored. Here we show that germacrene A oxidase (GAO), evolutionarily conserved in all major subfamilies of Asteraceae, catalyzes three consecutive oxidations of germacrene A to yield germacrene A acid. Furthermore, it is also capable of oxidizing non-natural substrate amorphadiene. Co-expression of lettuce GAO with germacrene synthase in engineered yeast synthesized aberrant products, costic acids and ilicic acid, in an acidic condition. However, cultivation in a neutral condition allowed the de novo synthesis of a single novel compound that was identified as germacrene A acid by gas and liquid chromatography and NMR analyses. To trace the evolutionary lineage of GAO in Asteraceae, homologous genes were further isolated from the representative species of three major subfamilies of Asteraceae (sunflower, chicory, and costus from Asteroideae, Cichorioideae, and Carduoideae, respectively) and also from the phylogenetically basal species, Barnadesia spinosa, from Barnadesioideae. The recombinant GAOs from these genes clearly showed germacrene A oxidase activities, suggesting that GAO activity is widely conserved in Asteraceae including the basal lineage. All GAOs could catalyze the three-step oxidation of non-natural substrate amorphadiene to artemisinic acid, whereas amorphadiene oxidase diverged from GAO displayed negligible activity for germacrene A oxidation. The observed amorphadiene oxidase activity in GAOs suggests that the catalytic plasticity is embedded in ancestral GAO enzymes that may contribute to the chemical and catalytic diversity in nature. 相似文献
4.
The climbing habit has evolved independently in many plant taxa, offering vines the ability to compete with non-climbing vegetation
for resources such as light, nutrients, and water. This review examines the structural and functional characteristics that
allow climbing plants to (1) achieve widespread dispersal, (2) transport large amounts of water throughout vessels, (3) maintain
high photosynthesis levels through a large leaf area to biomass ratio, (4) achieve rapid vertical and horizontal expansion
by fast growth rates and various climbing mechanisms and (5) survive and recover from disturbances. Due to the competitive
effects of vines on trees, management of vine growth is used to preserve tropical timber plantations, combat invasive weeds,
and promote rainforest recovery. In order to sustainably manage the vines into the future, it is necessary to understand the
mechanisms by which they can alter tropical forest succession and the impacts of various management techniques. 相似文献
5.
Marion S. Ecob Gillian S. Butler-Browne Robert G. Whalen 《Differentiation; research in biological diversity》1984,25(1-3):84-87
Abstract. Organotypic nerve-muscle cultures were prepared from foetal mouse spinal cord and adult mouse muscle fibres. In this system, the adult fibres degenerate and new myotubes form. The regenerated muscle fibres become innervated, develop cross-striations, and survive for several months. We have investigated the isozymes of myosin present in these muscle fibres using histochemistry and immunocytochemistry with antibodies to rat embryonic, neonatal, and adult fast myosins. We demonstrate that some of the regenerated fibres contain adult fast but not embryonic or neonatal myosin. This is the first demonstration of the production of adult myosin heavy chain in tissue culture. This system therefore offers the possibility for further study of the development of adult myosin isoforms in vitro. 相似文献
6.
Gillian D Alton David L Pearl Ken G Bateman W Bruce McNab Olaf Berke 《BMC veterinary research》2010,6(1):42
Background
Ontario provincial abattoirs have the potential to be important sources of syndromic surveillance data for emerging diseases of concern to animal health, public health and food safety. The objectives of this study were to: (1) describe provincially inspected abattoirs processing cattle in Ontario in terms of the number of abattoirs, the number of weeks abattoirs process cattle, geographical distribution, types of whole carcass condemnations reported, and the distance animals are shipped for slaughter; and (2) identify various seasonal, secular, disease and non-disease factors that might bias the results of quantitative methods, such as cluster detection methods, used for food animal syndromic surveillance. 相似文献7.
Rupert Cecil Gillian B. Robinson 《Biochimica et Biophysica Acta (BBA)/General Subjects》1975,404(1):164-168
It is known that insulin is adsorbed onto glass but it has been assumed that it is not adsorbed onto plastic. We find that tritium-labelled insulin is adsorbed onto all materials tried. The adsorption is reduced in the presence of other proteins and can be prevented altogether by coating the vessels with cetyl alcohol. 相似文献
8.
The rapid increase in protein synthesis that occurs on addition of insulin (1 mU/ml) to stepped-down 3T3 cells was blocked by pre-incubation of the cells with pertussis toxin. Cholera toxin on the other hand stimulated protein synthesis and this effect was insensitive to actinomycin D and inhibited by pro-treatment of the cells with phorbol dibutyrate to deplete cell protein kinase C. Insulin was found to cause a rapid and transient increase in diacylglycerol (DAG) synthesis. The insulin-induced increase in diacylglycerol was blocked by pertussis toxin. Exogenous DAG (10 M) stimulated protein synthesis within 1 hour. The results suggest that insuIin stimulates ribosomal activity through a signal mechanism that involves a G-protein mediated activation of phospholipase C to increase DAG levels. 相似文献
9.
P. L. Hope E. B. Cady A. Chu D. T. Delpy R. M. Gardiner E. O. R. Reynolds 《Journal of neurochemistry》1987,49(1):75-82
Brain metabolism and intracellular pH were studied during and after episodes of ischaemia and hypoxia-ischaemia in lambs anaesthetised with sodium pentobarbitone. 31P and 1H magnetic resonance spectroscopy methods were used to monitor brain pHi and brain concentrations of Pi, phosphocreatine (PCr), beta--nucleoside triphosphate (beta NTP), and lactate. Simultaneous measurements were made of cerebral blood flow and cerebral oxygen and glucose consumption. Cerebral ischaemia sufficient to reduce oxygen delivery to 75% of control values was associated with a fall in brain pHi and increase in brain Pi. Progressively severe hypoxia-ischaemia was associated with a progressive fall in brain pHi, PCr, and beta NTP and increase in brain Pi. In two animals the increase in brain lactate during hypoxia-ischaemia measured by 1H nuclear magnetic resonance (NMR) could be quantitatively accounted for by the increased net uptake of glucose by the brain in relation to oxygen, but was insufficient to account for the concomitant acidosis according to previous estimates of brain buffering capacity. In four animals brain pHi, PCr, Pi, and beta NTP had returned to normal 1 h after the hypoxic-ischaemic episode. In one animal brain pHi had reverted to normal at a time when 1H NMR indicated persistent elevation of brain lactate. 相似文献
10.
We have carried out the third in a series of anatomical dissections of amputated congenitally deformed human limbs in an attempt to determine the etiological relationship between the bony and soft tissue anomalies. This specimen consisted of a limb with congenital tibial aplasia and an adducted foot with five toes. The arterial and nerve patterns were reminiscent of those seen previously by us. The arteries were characterized by absence of the anterior tibial artery and an incomplete plantar arch. The superficial peroneal nerve terminated at the ankle. Extra branches from the sural, deep peroneal, and medial plantar nerves supplied branches to the dorsum of the foot. There were two extra muscles in the leg. The remaining muscles were normal, with the exception that most muscles normally inserting on the plantar surface of the foot inserted instead into a common tendon sheet. The tibia was replaced by a tendinous band, and the talus and calcaneus were united by a complete synchondrosis. The contralateral limb clinically appeared to have a clubfoot. The combination of an absent anterior tibial artery and an incomplete plantar arch is consistent with our theory that a reduced number of vessels puts the embryonic limb at increased risk of congenital defects due to the reduction in the number of collateral blood routes. Some event, such as extravasation of blood or embolization, may concurrently or subsequently compromise blood flow in the remaining vessels. We have previously observed abnormal arterial patterns similar to that described above in limbs having absence of the tibia and in other patients having clubfeet. The presence of abnormal arterial patterns in a limb with absence of the tibia and a contralateral limb with clubfoot suggests that absent tibia and clubfoot may be etiologically related. 相似文献