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Joseph A. Marsh 《Journal of molecular biology》2009,391(2):359-318
Obtaining detailed structural models of disordered states of proteins under nondenaturing conditions is important for a better understanding of both functional intrinsically disordered proteins and unfolded states of folded proteins. Extensive experimental characterization of the drk N-terminal SH3 domain unfolded state has shown that, although it appears to be highly disordered, it possesses significant nonrandom secondary and tertiary structure. In our previous attempts to generate structural models of the unfolded state using the program ENSEMBLE, we were limited by insufficient experimental restraints and conformational sampling. In this study, we have vastly expanded our experimental restraint set to include 1H-15N residual dipolar couplings, small-angle X-ray scattering measurements, nitroxide paramagnetic relaxation enhancements, O2-induced 13C paramagnetic shifts, hydrogen-exchange protection factors, and 15N R2 data, in addition to the previously used nuclear Overhauser effects, amino terminal Cu2+-Ni2+ binding paramagnetic relaxation enhancements, J-couplings, chemical shifts, hydrodynamic radius, and solvent accessibility restraints. We have also implemented a new ensemble calculation methodology that uses iterative conformational sampling and seeks to calculate the simplest possible ensemble models. As a result, we can now generate ensembles that are consistent with much larger experimental data sets than was previously possible. Although highly heterogeneous and having broad molecular size distributions, the calculated drk N-terminal SH3 domain unfolded-state ensembles have very different properties than expected for random or statistical coils and possess significant nonnative α-helical structure and both native-like and nonnative tertiary structure. 相似文献
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Megan F. Marchetti Matthew H. Persons 《Ethology : formerly Zeitschrift fur Tierpsychologie》2020,126(12):1111-1121
Life history theory predicts that iteroparous animals adaptively partition reproductive effort between current and future reproduction. When rearing costs of current offspring exceed the potential benefits, parental care should be terminated and deferred toward future reproduction. We tested two related predictions that follow from life history theory: (a) parents should be sensitive to offspring viability and withhold parental care if offspring survival probability drops and future reproductive opportunities are likely, and (b) parents should be less sensitive to offspring survival probability when future reproduction is unlikely and maximize parental care late in life. The wolf spider, Pardosa milvina, demonstrates extensive parental care; however, they may also abandon or cannibalize their egg sacs. We tested the effects of egg sac damage and production of a previous egg sac on egg sac abandonment and cannibalism decisions. Among four egg sac groups (1st egg sac intact, 1st egg sac damaged, 2nd egg sac intact, 2nd egg sac damaged), we daily monitored egg sac abandonment and cannibalism and measured differences in egg sac searching, protection, and grooming among removed and damaged egg sacs (N = 116 with 1st egg sac and 88 with 2nd egg sac). Females with first egg sacs abandoned and cannibalized damaged egg sacs significantly more compared to unmanipulated egg sacs; however, females with second egg sacs were insensitive to egg sac damage. Females also spent significantly more time protecting second egg sacs compared to first egg sacs and groomed damaged egg sacs significantly more than undamaged. These results support the general predictions of life history theory that indicate that abandonment and cannibalism should decrease with diminished future reproductive potential and that parents should be less sensitive to indicators of offspring survival probability late in life. 相似文献
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EB病毒LMP1-CTAR3对NP69细胞增殖和蛋白质表达的影响 总被引:1,自引:0,他引:1
为了探讨EB病毒潜伏性膜蛋白 1(LMP1)第三个功能活性区域(CTAR3)在鼻咽上皮细胞NP69中的转化作用机制,采用逆病毒感染的方法,将浓缩的逆病毒RV-LMP1和RV-LMP1Δ232~351分别感染鼻咽上皮细胞NP69,建立NP69-LMP1与NP69-LMP1Δ232~351稳定表达细胞系.通过绘制生长曲线、平皿克隆形成试验和软琼脂集落形成试验比较野生型和突变型LMP1对NP69细胞增殖的影响,运用蛋白质组学方法鉴定NP69-LMP1与NP69-LMP1Δ232~351细胞间的差异表达蛋白,选用实时荧光定量RT-PCR与Western blot对其中部分蛋白质点差异表达进行验证.结果发现:a.突变型LMP1Δ232~351促NP69细胞增殖的能力较野生型LMP1明显降低(n=3,P < 0.05);b.鉴定了LMP1-CTAR3在NP69细胞中参与调节的16个蛋白质(表达上调的蛋白质8个,下调的8个).c.实时荧光定量RT-PCR和Western blot证实了部分上述蛋白质的差异表达.以上结果说明,LMP1-CTAR3是其发挥促细胞增殖的重要活性部位,可能通过参与调节G蛋白和异柠檬酸脱氢酶等蛋白质的表达而起作用. 相似文献
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Activity-Dependent Bulk Synaptic Vesicle Endocytosis—A Fast,High Capacity Membrane Retrieval Mechanism 总被引:1,自引:0,他引:1
M. A. Cousin 《Molecular neurobiology》2009,39(3):185-189
Central nerve terminals are placed under considerable stress during intense stimulation due to large numbers of synaptic vesicles
(SVs) fusing with the plasma membrane. Classical clathrin-dependent SV endocytosis cannot correct for the large increase in
nerve terminal surface area in the short term, due to its slow kinetics and low capacity. During such intense stimulation,
an additional SV retrieval pathway is recruited called bulk endocytosis. Recent studies have shown that bulk endocytosis fulfils
all of the physiological requirements to remedy the acute changes in nerve terminal surface area to allow the nerve terminal
to continue to function. This review will summarise the recent developments in the field that characterise the physiology
of bulk endocytosis which show that it is a fast, activity-dependent and high capacity mechanism that is essential for the
function of central nerve terminals. 相似文献