A cytosolic iron chaperone that delivers iron to ferritin

Ferritins are the main iron storage proteins found in animals, plants, and bacteria. The capacity to store iron in ferritin is essential for life in mammals, but the mechanism by which cytosolic iron is delivered to ferritin is unknown. Human ferritins expressed in yeast contain little iron. Human poly (rC)-binding protein 1 (PCBP1) increased the amount of iron loaded into ferritin when expressed in yeast. PCBP1 bound to ferritin in vivo and bound iron and facilitated iron loading into ferritin in vitro. Depletion of PCBP1 in human cells inhibited ferritin iron loading and increased cytosolic iron pools. Thus, PCBP1 can function as a cytosolic iron chaperone in the delivery of iron to ferritin.     

N-terminal acetylation acts as an avidity enhancer within an interconnected multiprotein complex

Although many eukaryotic proteins are amino (N)-terminally acetylated, structural mechanisms by which N-terminal acetylation mediates protein interactions are largely unknown. Here, we found that N-terminal acetylation of the E2 enzyme, Ubc12, dictates distinctive E3-dependent ligation of the ubiquitin-like protein Nedd8 to Cul1. Structural, biochemical, biophysical, and genetic analyses revealed how complete burial of Ubc12's N-acetyl-methionine in a hydrophobic pocket in the E3, Dcn1, promotes cullin neddylation. The results suggest that the N-terminal acetyl both directs Ubc12's interactions with Dcn1 and prevents repulsion of a charged N terminus. Our data provide a link between acetylation and ubiquitin-like protein conjugation and define a mechanism for N-terminal acetylation-dependent recognition.     

The protein kinase p90 rsk as an essential mediator of cytostatic factor activity

Persistent activation of p42 mitogen-activated protein kinase (p42 MAPK) during mitosis induces a "cytostatic factor" arrest, the arrest responsible for preventing the parthenogenetic activation of unfertilized eggs. The protein kinase p90 Rsk is a substrate of p42 MAPK; thus, the role of p90 Rsk in p42 MAPK-induced mitotic arrest was examined. Xenopus laevis egg extracts immunodepleted of Rsk lost their capacity to undergo mitotic arrest in response to activation of the Mos-MEK-1-p42 MAPK cascade of protein kinases. Replenishing Rsk-depleted extracts with catalytically competent Rsk protein restored the ability of the extracts to undergo mitotic arrest. Rsk appears to be essential for cytostatic factor arrest.     

布氏田鼠肝脏内线粒体蛋白含量及细胞色素c氧化酶活性的变化

为探讨线粒体能量代谢中细胞色素c氧化酶(cytochromecoxidase,CcO)在布氏田鼠(Microtusbrandti)生长发育过程中的作用,应用紫外分光光度法测定了不同日龄的幼鼠肝脏内线粒体蛋白含量及CcO活性。结果发现根据幼鼠肝脏内线粒体蛋白的含量可将生长发育大致分为3个阶段:1～3日龄,4～19日龄,20日龄至成体。CcO活性变化趋势则呈倒钟形,以17日龄的CcO活性为最高。据此推测布氏田鼠肝脏内CcO的活性可能与恒温机制的建立有关系。     

具有抗氧化活性的猪血浆蛋白酶解产物的制备及理化特性

为系统考察酶解时间、酶种类和酶加入的顺序对猪血浆蛋白酶解产物抗氧化性的影响,采用风味蛋白酶、碱性蛋白酶及双酶(风味蛋白酶,碱性蛋白酶不同的加入顺序)在各自最适条件下对猪血浆蛋白进行酶解,并研究酶解产物的功能特性。结果表明:0~120min内,随着酶解时间的延长,水解度升高,各酶解产物的抗氧化性均有所增强;60~180min内,双酶酶解产物的清除DPPH自由基的能力、亚铁离子螯合能力和还原力显著高于单酶酶解产物(P0.05);90~150min内,先加碱性蛋白酶后加入风味蛋白酶酶解的酶解产物的水解度、清除DPPH自由基能力和还原力显著高于先加风味蛋白酶酶解后加入碱性蛋白酶酶解的酶解产物(P0.05);具有较高抗氧化性的酶解产物在不同pH条件下均有较好的溶解性和热稳定性;溶解性均在90%以上。     

Design and evolution of new catalytic activity with an existing protein scaffold

The design of enzymes with new functions and properties has long been a goal in protein engineering. Here, we report a strategy to change the catalytic activity of an existing protein scaffold. This was achieved by simultaneous incorporation and adjustment of functional elements through insertion, deletion, and substitution of several active site loops, followed by point mutations to fine-tune the activity. Using this approach, we were able to introduce beta-lactamase activity into the alphabeta/betaalpha metallohydrolase scaffold of glyoxalase II. The resulting enzyme, evMBL8 (evolved metallo beta-lactamase 8), completely lost its original activity and, instead, catalyzed the hydrolysis of cefotaxime with a (kcat/Km)app of 1.8 x 10(2) (mole/liter)(-1) second(-1), thus increasing resistance to Escherichia coli growth on cefotaxime by a factor of about 100.     

Pion cancer therapy: positron activity as an indicator of depth-dose

Negative pions are useful for radiation therapy of localized carcinoma because of their large end-of-range energy deposition upon capture in nuclei and the potentially accurate spatial localization of the radiation dose. Accurate determination of the depth of penetration of the pion beam in vivo can be accomplished by counting the back-to-back annihilation gamma rays resulting from positron activity induced by the stopped pions.     

应用线粒体基因序列分析虎的进化关系

【目的】研究华南虎Panthera tigris amoyensis、东北虎P.tigris altaica和孟加拉虎P.tigris tigris之间的进化关系.【方法】采用PCR扩增结合质粒测序的方法,获得东北虎、孟加拉虎和华南虎线粒体DNA COⅠ基因750 bp序列、COⅢ基因737 bp的序列和ND4基因561 bp的序列.使用DNAStar和DNAssit软件对这些序列进行碱基组成、序列间的转换/颠换值、遗传距离等分析.选择家猫Felis catus、猎豹Acinonyx jubatus及云豹Neofelis nebulosa的mtDNA作为外群,利用Mega 4.0采用邻接法重建华南虎、东北虎、孟加拉虎的进化关系树.【结果和结论】研究发现东北虎、孟加拉虎和华南虎线粒体DNA COⅠ、COⅢ和ND4基因变异位点较少,且华南虎同孟加拉虎、东北虎的遗传距离要大于孟加拉虎和东北虎的遗传距离;构建了华南虎、东北虎、孟加拉虎的COⅠ、COⅢ和ND4基因的基因树,通过对基因树分析可以得东北虎和孟加拉均是由华南虎进化而来的结论.     

Computer graphics as an aid to learning

Departing from the traditional model for teaching and learning, this article deals with the problem of teaching and learning the effective application of knowledge already acquired. To this end, a model for the process of exercising judgment was outlined, and the results of an empirical study of judgmental learning were employed to show the inadequacy of the traditional outcome feedback procedures. Computer graphics techniques were used to provide new forms of information to the learner; the results are promising for the rapid learning of a task that would otherwise be difficult to learn.     

A 13-kilodalton maize mitochondrial protein in E. coli confers sensitivity to Bipolaris maydis toxin

The Texas male-sterile cytoplasm (cms-T) of maize carries the cytoplasmically inherited trait of male sterility. Mitochondria isolated from cms-T maize are specifically sensitive to a toxin (BmT-toxin) produced by the fungal pathogen Bipolaris maydis, race T, and the carbamate insecticide methomyl. A mitochondrial gene unique to cms-T maize, which produces a 13-kilodalton polypeptide associated with cytoplasmic male sterility, was expressed in Escherichia coli. After addition of BmT-toxin or methomyl, inhibition of whole cell respiration and swelling of spheroplasts were observed in Escherichia coli cultures producing the novel mitochondrial protein; these effects are similar to those observed with isolated cms-T mitochondria. The amino-terminal region of the 13-kilodalton polypeptide appears to be essential for proper interaction with the BmT-toxin and methomyl. These results implicate the 13-kilodalton polypeptide in conferring toxin sensitivity to mitochondria of cms-T maize.     

Back-action evasion as an alternative to impedance matching

Back-action evasion is a measurement technique originally devised to overcome certain limits imposed by quantum mechanics on the sensitivity of gravitational radiation detectors. The technique is, however, more generally applicable and can be used to improve the sensitivity of instrumentation with noise floors much greater than the quantum noise floor. The principle of back-action evasion is described here by means of a simple example. A comparison of back-action evasion with impedance matching is made to clarify when back-action evasion may be useful. Back-action evasion allows one to achieve a sensitivity comparable to that achieved by impedance matching.     

Hemolin: an insect-immune protein belonging to the immunoglobulin superfamily

Insects have an efficient defense system against infections. Their antibacterial immune proteins have been well characterized. However, the molecular mechanisms by which insects recognize foreignness are not yet known. Data are presented showing that hemolin (previously named P4), a bacteria-inducible hemolymph protein of the giant silk moth Hyalophora cecropia, belongs to the immunoglobulin superfamily. Functional analyses indicate that hemolin is one of the first hemolymph components to bind to the bacterial surface, taking part in a protein complex formation that is likely to initiate the immune response.     

极细链格孢菌66 kDa激活蛋白的纯化及诱导烟草抗TMV的功能

激活蛋白是一类来源于真菌、诱导植物获得系统抗性的蛋白激发子。通过加热离心、乙醇沉淀、离子交换层析、超滤等方法,从极细链格孢菌(Alternaria tenuissi ma)JH505菌株的菌丝体中分离纯化出一种激活蛋白,在SDS-PAGE银染图谱显示单一条带,表观分子量约为66 kDa,糖蛋白特异性染色确定其为糖蛋白。该蛋白可以诱导三生烟提高对烟草花叶病毒(Tobacco mosaic virus,TMV)的抗性,处理后的三生烟叶片TMV枯斑数显著减少,枯斑抑制率可达到49%。     

Conversion of a PI-anchored protein to an integral membrane protein by a single amino acid mutation

Qa-2, a cell-surface glycoprotein anchored by phosphatidylinositol (PI), is structurally related to the class I transplantation antigens H-2 K, D, and L, which are integral membrane glycoproteins. The predicted transmembrane segment of Qa-2 differs from those of H-2 K, D, and L by the presence of an aspartate in place of a valine at position 295. A single base change that replaced this aspartate with valine resulted in cell-surface Qa-2 molecules that were insensitive to hydrolysis by a PI-specific phospholipase C and more resistant to papain cleavage, properties shared by H-2D. Cells expressing Asp----Val mutant Qa-2 proteins were still able to attach a PI anchor to endogenous proteins such as Thy-1 and J11D. It therefore appears that this single amino acid change converts Qa-2 from a PI-linked form into an integral membrane protein.     

传染性法氏囊病毒VP2蛋白的表达及间接 ELISA抗体检测方法的建立

建立以传染性法氏囊病毒(IBDV)VP2蛋白为诊断抗原的传染性法氏囊病(IBD)血清学检测方法,研究了VP2蛋白的编码基因,并在原核表达系统中表达了VP2蛋白。用表达的VP2蛋白建立了IBDV间接ELISA检测方法。抗原最佳包被量为每孔174.67 ng,血清最佳稀释度为1:40。对采自安徽部分地区的979鸡血清样品进行检测,IBDV抗体阳性率为39.73%。结果证实,间接ELISA法用于IBD血清抗体的监测具有良好的特异性,是一种快速简便的血清学方法,值得在基层推广应用。     

Ubiquitin protein ligase activity of IAPs and their degradation in proteasomes in response to apoptotic stimuli

To determine why proteasome inhibitors prevent thymocyte death, we examined whether proteasomes degrade anti-apoptotic molecules in cells induced to undergo apoptosis. The c-IAP1 and XIAP inhibitors of apoptosis were selectively lost in glucocorticoid- or etoposide-treated thymocytes in a proteasome-dependent manner before death. IAPs catalyzed their own ubiquitination in vitro, an activity requiring the RING domain. Overexpressed wild-type c-IAP1, but not a RING domain mutant, was spontaneously ubiquitinated and degraded, and stably expressed XIAP lacking the RING domain was relatively resistant to apoptosis-induced degradation and, correspondingly, more effective at preventing apoptosis than wild-type XIAP. Autoubiquitination and degradation of IAPs may be a key event in the apoptotic program.     

Bivoltinism as an antecedent to eusociality in the paper wasp genus Polistes

To learn the evolutionary trajectories of caste differentiation in eusocial species is a major goal of sociobiology. We present an explanatory framework for caste evolution in the eusocial wasp genus Polistes (Vespidae), which is a model system for insect eusocial evolution. We hypothesize that Polistes worker and gyne castes stem from two developmental pathways that characterized the bivoltine life cycle of a solitary ancestor. Through individual-based simulations, we show that our mechanistic framework can reproduce colony-level characteristics of Polistes and, thereby, that social castes can emerge from solitary regulatory pathways. Our explanatory framework illustrates, by specific example, a changed perspective for understanding insect social evolution.