Proteomic analysis of differentially expressed proteins in lymphoid organ of Fenneropenaeus chinensis response to Vibrio anguillarum stimulation

To gain an insight into the function of shrimp lymphoid organ at protein level, we analyzed the proteome of lymphoid organ in healthy Chinese shrimp Fenneropenaeus chinensis (F. chinensis) through two-dimensional gel electrophoresis (2-DE) based proteomic approach. A total of 95 spots representing 75 protein entries were identified by liquid chromatography tandem mass spectrometry (LC-MS/MS) with both online and in-house database. According to Gene Ontology (GO) annotation of biological process, the identified proteins were classified into 13 categories. Among them, approximately 36% of proteins related to cytoskeleton are noticeable. Then, a comparative proteomic approach was employed to investigate the differentially expressed proteins in lymphoid organ of Vibrio anguillarum-challenged F. chinensis. At 24 h post-injection (hpi), 17 differentially expressed protein spots were successfully identified, including 4 up-regulated protein spots (represent 4 proteins: cathepsin L protein similar to squid CG16901-PC, protein kinase C and protein similar to T-complex Chaperonin 5 CG8439-PA), and 13 down-regulated protein spots (represent 9 proteins: actin, beta-actin, cytoplasmic actin CyII, alpha tubulin, beta tubulin, protein similar to proteasome delta, vacuolar ATP synthase subunit B, elongation factor 2, carboxypeptidase B). These data may help us to understand the function of lymphoid organ and the molecular immune mechanism of shrimp responsive to pathogen infection. (C) 2010 Elsevier Ltd. All rights reserved.

三种红藻光合作用色素系统的比较研究

本研究分为三个部分：1.以坛紫菜（Porphyra haitanesis Chang et Zheng）的叶状体和丝状体为研究对象，比较坛紫菜叶状体和丝状体的光合色素、色素蛋白的组成，并提取纯化藻红蛋白、藻蓝蛋白、藻胆体及类囊体膜和光系统。研究结果表明坛紫菜叶状体和丝状体色素及色素蛋白的含量不同，藻红蛋白是主要的色素蛋白，坛紫菜叶状体和丝状体的藻红蛋白的含量分别为2.9mg藻红蛋白/g鲜重、4.2mg藻红蛋白/g鲜重，这表明坛紫菜叶状体和丝状体藻红蛋白含量丰富，是提取藻红蛋白很好的材料。藻胆体的性质差异不大，但类囊体膜差异显著，从坛紫菜叶状体中分离到了两种不同的类囊体膜带，光系统Ⅰ(PSⅠ)和PSⅡ分别结合在两条类囊体膜带上，但从坛紫菜丝状体中也分离到两条类囊体膜带，它们的光谱性质和蛋白组成相似，仅放氧速率和DCIP活性有差异，从坛紫菜丝状体中我们仅分离到PSⅡ。坛紫菜叶状体PSⅡ有5种外在蛋白(33、20、Cytc 550、15、12kDa蛋白)，而坛紫菜丝状体外在蛋白仅有4条，缺少12kDa蛋白。2. 以在中国江苏部分地区进行了大规模的商业化栽培的突变体条斑紫菜（Porphyra yezoensis Ueda）和野生型条斑紫菜为研究对象，比较其色素及色素蛋白组成、对不能光质的利用率及藻胆体的组成。条斑紫菜和突变型条斑紫菜对不同的光质利用效果有差异，在白光的照射下，野生型紫菜的放氧速率最大，而突变型紫菜在黄光照射下的放氧速率最大。条斑紫菜野生型与突变型色素含量上有明显的差异,突变型紫菜的藻红蛋白含量明显减少而藻蓝蛋白的含量增加。通过杂交的方法证实诱变所获得条斑紫菜突变体为细胞质突变，但是突变型紫菜却发生了由细胞核编码的γ亚基的缺失，这表明突变型紫菜藻红蛋白含量和性质发生了明显的变化。3. 为了找出淡水红藻-深紫美芒藻(Compsopogon coeruleus (Balbis) Montagne)分布狭窄及生物产量低的原因，本文对深紫美芒藻在不同的盐离子浓度下的放氧速率及藻胆体色素组成和结构上进行研究。结果显示：微量的NaCl（0.1mM）促进深紫美芒藻放氧，而深紫美芒藻在较高的NaCl（1、10mM）, NaH2PO4 （0.1、1、10mM）和 NH4NO3（0.1、1、10mM）溶液中却没有检测到氧气的产生。这与深紫美芒藻生长的环境一致即深紫美芒藻生活在低盐浓度、低营养的泉水中。深紫美芒藻的藻胆体是由藻红蛋白、藻蓝蛋白及别藻蓝蛋白组成，上面结合α、β和γ亚基，含有藻红胆素、藻篮胆素，但缺乏缺少藻尿胆素。

栉孔扇贝大规模死亡病原学研究

用平板画线法从患病栉孔扇贝（Chlamys farreri）体内分离到了一种原核生物（简称QDP）。QDP可以在改进的液体培养基MEM（含2.2%NaCl，5%小牛血清）和脑心浸液（含2.2% NaCl）中生长；菌落在显微镜下（150×）为无色、透明的小点状；革兰氏染色阴性；菌体为圆形或近似圆形。QDP在发育过程中有两种状态，一种为未成熟阶段，直径小于100nm；另一种为成熟阶段，直径变化很大，最小约60nm，最大可达4µm以上。较小的个体有拟核、核糖体和新月状的空泡，未见细胞壁；较大的个体有细胞壁，胞内大部分被空泡充满，未见拟核和核糖体。栉孔扇贝组织超簿切片电镜观查证实QDP的存在。QDP的密度随着生长发育时间的不同而有所变化，繁殖高峰期密度较大。 建立了密度梯度离心结合滤膜过滤分离技术，优化人工培养条件。最适生长温度为23℃，最适生长pH值为7.4，最适生长盐度相当于细胞培养液所需的盐浓度（0.85%NaCl）。 提取的QDP核酸能被RNase A 降解，且没有检测到DNA。以PCR、RT-PCR扩增其16SrRNA基因序列片段，PCR反应没有扩增出扩增子，而RT-PCR则扩增出了16S rRNA基因序列片段，经测定其序列全长度为1430bp，经与GENEBANK中的16S rRNA片段比较分析，与6种不同科的微生物的同源率最高的为95%-95.47%。 采用温度梯度和病原浓度梯度回归感染实验方法，较为系统地研究了QDP的致病性。研究结果表明：QDP对栉孔扇贝有强烈的致病作用，高温（23℃以上）是其致病的必要条件，证实DQP是栉孔扇贝大规模死亡的病原体之一。

Identification and mapping of amplified fragment length polymorphism markers linked to shell color in bay scallop, Argopecten irradians irradians (Lamarck, 1819)

Amplified fragment length polymorphisms (AFLP) were used to study the inheritance of shell color in Argopecten irradians. Two scallops, one with orange and the other with white shells, were used as parents to produce four F-1 families by selfing and outcrossing. Eighty-eight progeny, 37 orange and 51 white, were randomly selected from one of the families for segregation and mapping analysis with AFLP and microsatellite markers. Twenty-five AFLP primer pairs were screened, yielding 1138 fragments, among which 148 (13.0%) were polymorphic in two parents and segregated in progeny. Six AFLP markers showed significant (P < 0.05) association with shell color. All six loci were mapped to one linkage group. One of the markers, F1f335, is completely linked to the gene for orange shell, which we designated as Orange1, without any recombination in the progeny we sampled. The marker was amplified in the orange parent and all orange progeny, but absent in the white parent and all the white progeny. The close linkage between F1f335 and Orange1 was validated using bulk segregation analysis in two natural populations, and all our data indicate that F1f335 is specific for the shell color gene, Orange1. The genomic mapping of a shell color gene in bay scallop improves our understanding of shell color inheritance and may contribute to the breeding of molluscs with desired shell colors.

基于有限状态机的空间对接仿真平台控制系统设计

给出了一种基于分层式有限状态机的五自由度空间对接仿真平台控制系统设计方法,对状态机进行了扩展定义.增加了一个定义于状态上的变量属性集合,使其有利于系统的代码实现.结合控制系统采用的10 ms定时中断机制,将状态机层次划分到了可以分析每个10 ms硬件中断程序所需实现的控制功能状态及其转移、继承关系的程度,可以更清晰地设计出中断程序所需要的构成结构.系统的实际应用结果证明了上述方法的有效性.

大别山岩石圈有效弹性厚度初步研究

As powerful tools to study the lithosphere dynamics, the effective elastic thickness (Te) as well as the envelope of yielding stress of lithosphere have been attracted great attention of geoscientists in the past thirty years. The oceanic lithosphere, contrary to the continental lithosphere, has more fruits for its simple structures and evolution process. In continent, the lithosphere commonly is complex and variable in the rheological, thermal structures, and has a complicated history. Therefore, the application of the effective elastic thickness in continent is still a subject to learn in a long time. Te, with the definition of the thickness of an elastic plate in theory flexured by the equal benging of the real stress in the lithosphere plate (Turcotte, 1982), marks the depth of transition between elastic and fluid behaviors of rocks subjected to stress exceeding 100 MPa over the geological timescales (McNutt, 1990). There are three methods often adapted: admittance or isostatic response function, coherence and forwarding. In principle, the models of Te consist of thermal-rheological, non-linear Maxwell, non-linear work hardening and rheological layered models. There is a tentative knowledge of Te that it is affected by the following factors: crustal thickness, crust-mantle decoupling, plate bending, boundary conditions of plate (end forces and bending moments), stress state, sedimentary layer, faulting effect, variation in the mountain belts' strike, foreland basin, inheritance of tectonic evolution, convection of mantle, seismic depth and lithosphere strength. In this thesis, the author introduces the geological sketch of the Dabie collisional orogenic belt and the Hefei Basin. The Dabie Mts. is famous for the ultra-high pressure metamorphism. The crustal materials subducted down to the depth of at least 100 km and exhumed. So that the front subjects arise such as the deeply subduction of continent, and the post-collisional crust-mantle interaction. In a geological journey at June of 1999, the author found the rarely variolitic basaltic andesite in the Dabie Mts. It occurs in Susong Group, near Zhifenghe Countryside, Susong County, Anhui Province. It is just to the south of the boundary between the high-grade Susong melange and the ultra-high grade South Dabie melange. It has a noticeable knobby or pitted appearance in the surface. The size of the varioles is about 1-4 mm. In hand-specimen and under microscope, there are distinct contacts between the varioles and the matrice. The mineralogy of the varioles is primarily radiate plagioclase, with little pyroxene, hornblende and quartz. The pyroxene, hornblende and quartz are in the interstices between plagioclase. The matrix is consisted of glass, and micro-crystals of chlorite, epidote and zoisite. It is clearly subjected and extensive alteration. The andesite has an uncommon chemical composition. The SiO_2 content is about 56.8%, TiO_2 = 0.9%, MgO = 6.4%, (Fe_2O_3)_(Total) = 6.7% ～ 7.6%, 100 Mg/(Mg+Fe) = 64.1 ～ 66.2. Mg# is significantly high. The andesite has higher abundances of large-lithophile trace elements (e.g. K, Ba, Sr, LREE), e.g. La/Nd = 5.56-6.07, low abundances of high-strength-field elements (HFSE, e.g. Ta, Nb, P, Ti), particularly Ta and Nb strongly depleted. These are consistent with the characteristics of subducted-related magmas (Pearcce, 1982; Sun and McDonaugh, 1989). In the spider diagram of trace elements, from Ce to right hand, the abundances of elements decrease quickly, showing a characteristic of the continental margins (Pearce, 1982). There has a strongly enrichment of light-rare-earth elements, with a significant diffraction of REEs (the mean value of (La/Yb)_N is 32.84). No Eu anomaly, but there are anomaly high (La/Yb)_N = 28.63-36.74, (La/Y)_N = 70.33 - 82.84. The elements Y and Yb depleted greatly, Y < 20 ppm, Y_N = 2.74-2.84, Yb_N = 2.18 - 2.35. From the La-(La/Sm) diagram, the andesite is derived from partial melting. But the epsilone value of Nd is -18.7 ～ -19.2, so that the material source may be the mantle materials affected by the crustal materials. The Nd model age is 1.9 Ga indicating that the basaltic andesite was resulted from the post-collisional crust-mantle interaction between the subducted Yangze carton and the mantle of Sino-Korea carton. To obtain the Te of the lithosphere beneath the Dabie Mts. and the Hefei Basin, the author applies the coherence method in this thesis. The author makes two topography-gravity profiles (profiles 7~(th) and 9~(th)) across the Dabie Mts. and the Hefei Basin, and calculates the auto-coherence, across coherence, power spectrum, across power spectrum of the topography and gravity of the two profiles. From the relationships between the coherence and the wave-number of profiles. From the relationships between the coherence and the wave-number of profiles 7~(th) and 9~(th), it is obtained that the characteristic wavelengths respectively are 157 km and 126 km. Consequently the values of effective elastic thickness are 6.5 km and 4.8 km, respectively. However, the Te values merely are the minimum value of the lithosphere because the coherencemethod in a relative small region will generate a systemic underestimation. Why there is a so low Te value? In order to check the strength of the lithosphere beneath the Dabie Mts., the authore tries to outline the yielding-stress envelope of the lithosphere. It is suggested that the elastic layers in the crust and upper mantle are 18 km and 35 km, respectively. Since there exist a low viscosity layer about 3-5 km thickness, so it is reasonable that the decoupling between the crust and mantle occurred. So the effective thickness of the lithosphere can be estimated from the two elastic layers. Te is about 34 km. This is the maximum strength of the lithosphere. We can make an approximately estimation about the strength of the lithosphere beneath the Dabie Mts.: Te is about 20-30 km. The author believes that the following factors should be responsible for the low Te value: (1) the Dabie Mts. has elevated strongly since K_3-J_1. The north part of the Dabie Mts. elevates faster than the south part today; (2) there occur large active striking faults in this area. And in the east, the huge Tan-Lu striking fault anyway tends to decrease the lithosphere strength; (3) the lithosphere beneath the Dabie Mts. is heter-homogeneous in spatio-temporal; (4) the study area just locates in the adjacent region between the eastern China where the lithosphere thickness is significantly reduced and the normal western China. These factors will decrease the lithosphere strength.

Protocolo de tecnologia: selecao para resistencia a doencas em hortalicas. 1. Etapas para selecao de fontes de resistencia a doencas.

1999

Exploitation of Ebola Virus VP35 Protein to identify new drugs counteracting its type I IFN antagonism

Ebolaviruses (EBOVs) are among the most virulent and deadly pathogens ever known, causing fulminant haemorrhagic fevers in humans and non-human primates. The 2014 outbreak of Ebola virus disease (EVD) in West Africa has claimed more lives than all previous EVD outbreaks combined. The EBOV high mortality rates have been related to the virus-induced impairment of the host innate immunity reaction due to two virus-coded proteins, VP24 and VP35. EBOV VP35 is a multifunctional protein, it is essential for viral replication as a component of the viral RNA polymerase and it also participates in nucleocapsid assembly. Early during EBOV infection, alpha-beta interferon (IFN-α/β) production would be triggered upon recognition of viral dsRNA products by cytoplasmic retinoic acid-inducible gene I (RIG-I)-like receptors (RLRs). However, this recognition is efficiently prevented by the double-stranded RNA (dsRNA) binding activity of the EBOV VP35 protein, which hides RLRs binding sites on the dsRNA phosphate backbone as well the 5’-triphosphate (5’-ppp) dsRNA ends to RIG-I recognition. In addition to dsRNA binding and sequestration, EBOV VP35 inhibits IFN-α/β production preventing the activation of the IFN regulatory factor 3 (IRF-3) by direct interaction with cellular proteins. Previous studies demonstrated that single amino acid changes in the VP35 dsRNA binding domain reduce EBOV virulence, indicating that VP35 is an attractive target for antiviral drugs development. Within this context, here we report the establishment of a novel method to characterize the EBOV VP35 inhibitory function of the dsRNA-dependent RIG-I-mediated IFN-β signaling pathway in a BLS2 cell culture setting. In such system, a plasmid containing the promoter region of IFN-β gene linked with a luciferase reporter gene was transfected, together with a EBOV VP35 mammalian expression plasmid, into the IFN-sensitive A549 cell line, and the IFN-induction was stimulated through dsRNA transfection. Through alanine scanning mutational studies with biochemical, cellular and computational methods we highlighted the importance of some VP35 residues involved in dsRNA end-capping binding, such as R312, K282 and R322, that may serve as target for the development of small-molecule inhibitors against EBOV. Furthermore, we identified a synthetic compound that increased IFN-induction only under antiviral response stimulation and subverted VP35 inhibition, proving to be very attractive for the development of an antiviral drug. In conclusion, our results provide the establishment of a new assay as a straightforward tool for the screening of antiviral compounds that target i) dsRNA-VP35 or cellular protein-VP35 interaction and ii) dsRNA-dependent RIG-I-mediated IFN signaling pathway, in order to potentiate the IFN response against VP35 inhibition, setting the bases for further drug development.

Ruolo e limiti del contratto negli assetti familiari in funzione successoria

The Article 457 c.c. expressly excludes the contract by the sources of the succession. Moreover, the article 458 c.c., apart from the initial brief aside dedicated to the institute of the family pact, agrees nullity of the agreements with whom someone decides its own succession as well as those with which the future successor could decide about his rights or renounce to them about a succession not yet open. However, for a long time, the Italian doctrine wonders about the role of the contract within the succession law. It feels, in fact, the need to expand the private autonomy within the inheritance that is excessively sacrificed by the prohibition of succession agreements and by the norms for the protection of legitimate heirs. The reasons which led the legal science to these conclusions are based on different events, both social and economic, that push the interpreter to a modernization of dogmatic categories with which he can represent the succession mortis causa. In addiction, it is necessary to underline the crisis of the agreements mortis causa due to this economical and social events: as a matter of fact, the will, as the only way to give the assets post mortem, revealed itself incomplete and extremely severe compared to the new social needs. In fact, increasingly the way to give the assets happens out of the inheritance and despite to the institutions designed by the law. For these reasons, in order to adapt the system of succession to modern economic and social needs, the doctrine has identified, within the system, institutions of a contractual nature in order to better achieve the interests of private, obviating the limits assigned to the shop last will. And recently, in this context, our legislator has introduced the institution of the family pact (art. 768 bis et seq. c.c.), that is the agreement through “the entrepreneur transfers, in whole or in part , the company, and the holder of equity investments transfers, in whole or in part, its shares, to one or more descendants". While, however, part of the doctrine encourages the provision of tools that enable a person to have in advance of his succession, on the other hand there are those who promote the centrality of the will within our legal system and calls for the revitalization in respect of its vast potential is not always adequately exploited. This research aims to verify whether the contract can find importance within the phenomenon of succession for the inter vivos transfer of family assets and if the same has the characteristics to be considered a working alternative to the will. In the present work will be analyzed, in addition, some of the institutions that the doctrine has considered alternatives to the will and particularly the institution of the family pact. The survey will also be directed to the limits that the private autonomy and the legislator met in the use of the contractual instrument, limits that are mostly originated by the rules and principles of the law of succession.

Diurnal variation in uptake and xylem contents of inorganic and assimilated N under continuous and interrupted N supply to Phleum pratense and Festuca pratensis

J. H. Macduff and A. K. Bakken. (2003). Diurnal variation in uptake and xylem contents of inorganic and assimilated N under continuous and interrupted N supply to Phleum pratense and Festuca pratensis. Journal of Experimental Botany, 54 (381) pp.431-444 Sponsorship: BBSRC / Norwegian Crop Research Institute RAE2008

A Memory Model for Concept Hierarchy Representation and Commonsense Reasoning

Most associative memory models perform one level mapping between predefined sets of input and output patterns1 and are unable to represent hierarchical knowledge. Complex AI systems allow hierarchical representation of concepts, but generally do not have learning capabilities. In this paper, a memory model is proposed which forms concept hierarchy by learning sample relations between concepts. All concepts are represented in a concept layer. Relations between a concept and its defining lower level concepts, are chunked as cognitive codes represented in a coding layer. By updating memory contents in the concept layer through code firing in the coding layer, the system is able to perform an important class of commonsense reasoning, namely recognition and inheritance.

Osmotic stress tolerance mechanisms in Lactococcus lactis

The response of Lactococcus lactis subsp. cremoris NCDO 712 to low water activity (aw) was investigated, both in relation to growth following moderate reductions in the aw and in terms of survival following substantial reduction of the aw with NaCI. Lc.lactis NCDO 712 was capable of growth in the presence of ≤ 4% w/v NaCI and concentrations in excess of 4% w/v were lethal to the cells. The presence of magnesium ions significantly increased the resistance of NCDO 712 to challenge with NaCI and also to challenge with high temperature or low pH. Survival of Lc.lactis NCDO 712 exposed to high NaCI concentrations was growth phase dependent and cells were most sensitive in the early exponential phase of growth. Pre-exposure to 3% w/v NaCI induced limited protection against subsequent challenge with higher NaCI concentrations. The induction was inhibited by chloramphenicol and even when induced, the response did not protect against NaCI concentrations> 10% w/v. When growing at low aw, potassium was accumulated by Lc. lactis NCDO 712 growing at low aw, if the aw was reduced by glucose or fructose, but not by NaCI. Reducing the potassium concentration of chemically defined medium from 20 to 0.5 mM) produced a substantial reduction in the growth rate, if the aw was reduced with NaCI, but not with glucose or fructose. The reduction of the growth rate correlated strongly with a reduction in the cytoplasmic potassium concentration and in cell volume. Addition of the compatible solute glycine betaine, partially reversed the inhibition of growth rate and partially restored the cell volume. The potassium transport system was characterised in cells grown in medium at both high and low aw. It appeared that a single system was present, which was induced approximately two-fold by growth at low aw. Potassium transport was assayed in vitro using cells depleted of potassium; the assay was competitively inhibited by Na+ and by the other monovalent cations NH4+, Li+, and Cs+. There was a strong correlation between the ability of strains of Lc. lactis subsp. lactis and subsp. cremoris to grow at low aw and their ability to accumulate the compatible solute glycine betaine. The Lc. lactis subsp. cremoris strains incapable of growth at NaCI concentrations> 2% w/v did not accumulate glycine betaine when growing at low aw, whereas strains capable of growth at NaCI concentrations up to 4% w/v did. A mutant, extremely sensitive to low aw was isolated from the parent strain Lc. lactis subsp. cremoris MG 1363, a plasmid free derivative of NCDO 712. The parent strain tolerated up to 4% w/v NaCI and actively accumulated glycine betaine when challenged at low aw. The mutant had lost the ability to accumulate glycine betaine and was incapable of growth at NaCI concentrations >2% w/v or the equivalent concentration of glucose. As no other compatible solute seemed capable of substitution for glycine betaine, the data suggest that the traditional; phenotypic speciation of strains on the basis of tolerance to 4% w/v NaCI can be explained as possession or lack of a glycine betaine transport system.

Investigation of the genetic susceptibility to osteoporosis in the Irish population

Osteoporosis is a complex skeletal disorder characterized by compromised bone strength. Variation in bone mineral density (BMD) is a contributing factor. The aim of this research as to select informative single nucleotide polymorphisms (SNPs) in potential candidate genes from loci suggestively linked to BMD variation for fine mapping. The gene regulated by oestrogen in breast cancer 1 (GREB1), located at 2p25.1, was selected. GREB1 transcription is initiated early in the oestrogen receptor alpha regulated pathway. There was significant association between GREB1_03 and BMD variation at the lumbar spine and femoral neck (FN) in the discovery cohort. Significant association was observed between GREB1_04 and FN BMD in the replication cohort. The development and differentiation enhancing factor 2, the integrin cytoplasmic domain associated protein 1 and A-disintegrin and metalloprotease 17 were selected due to their respective roles in cell mobility and adhesion. There was no linkage or association observed between the Chr2 cluster SNPs and BMD. Two factors in bone remodelling are the attraction of bone cell precursors and endocrine regulation of the process, primarily through the action of parathyroid hormone (PTH). The C-C chemokine receptor type 3 (CCR3) encodes a CC chemokine receptor expressed in osteoclast precursors. The PTH receptor type 1 (PTHR1) encodes a G-protein coupled receptor for PTH. Association was observed between CCR3 haplotypes and BMD variation at the FN. There was no linkage or association observed between PTHR1 SNPs and BMD variation. Population genetic studies with complex phenotypes endeavour to elucidate the traits genetic architecture. This study presents evidence of association between GREB1 and BMD variation and as such, introduces GREB1 as a novel gene target for osteoporosis genetics studies. It affirms that common genomic variants in PTHR1 are not associated with BMD variation in Caucasians and supports the evidence that CCR3 may be contributing to BMD variation

The Islamic commercial crisis: Institutional roots of economic underdevelopment in the Middle East

During the second millennium, the Middle East's commerce with Western Europe fell increasingly under European domination. Two factors played critical roles. First, the Islamic inheritance system, by raising the costs of dissolving a partnership following a partner's death, kept Middle Eastern commercial enterprises small and ephemeral. Second, certain European inheritance systems facilitated large and durable partnerships by reducing the likelihood of premature dissolution. The upshot is that European enterprises grew larger than those of the Islamic world. Moreover, while ever larger enterprises propelled further organizational transformations in Europe, persistently small enterprises inhibited economic modernization in the Middle East.

HDAC6 and Ubp-M BUZ domains recognize specific C-terminal sequences of proteins.

The BUZ/Znf-UBP domain is a protein module found in the cytoplasmic deacetylase HDAC6, E3 ubiquitin ligase BRAP2/IMP, and a subfamily of ubiquitin-specific proteases. Although several BUZ domains have been shown to bind ubiquitin with high affinity by recognizing its C-terminal sequence (RLRGG-COOH), it is currently unknown whether the interaction is sequence-specific or whether the BUZ domains are capable of binding to proteins other than ubiquitin. In this work, the BUZ domains of HDAC6 and Ubp-M were subjected to screening against a one-bead-one-compound (OBOC) peptide library that exhibited random peptide sequences with free C-termini. Sequence analysis of the selected binding peptides as well as alanine scanning studies revealed that the BUZ domains require a C-terminal Gly-Gly motif for binding. At the more N-terminal positions, the two BUZ domains have distinct sequence specificities, allowing them to bind to different peptides and/or proteins. A database search of the human proteome on the basis of the BUZ domain specificities identified 11 and 24 potential partner proteins for Ubp-M and HDAC6 BUZ domains, respectively. Peptides corresponding to the C-terminal sequences of four of the predicted binding partners (FBXO11, histone H4, PTOV1, and FAT10) were synthesized and tested for binding to the BUZ domains by fluorescence polarization. All four peptides bound to the HDAC6 BUZ domain with low micromolar K(D) values and less tightly to the Ubp-M BUZ domain. Finally, in vitro pull-down assays showed that the Ubp-M BUZ domain was capable of binding to the histone H3-histone H4 tetramer protein complex. Our results suggest that BUZ domains are sequence-specific protein-binding modules, with each BUZ domain potentially binding to a different subset of proteins.