Peptide-protein interaction markedly alters the functional properties of the catalytic subunit of aspartate transcarbamoylase.

Interaction of a 70-amino acid zinc-binding polypeptide from the regulatory chain of aspartate transcarbamoylase (ATCase) with the catalytic (C) subunit leads to dramatic changes in enzyme activity and affinity for ligand binding at the active sites. The complex between the polypeptide (zinc domain) and wild-type C trimer exhibits hyperbolic kinetics in contrast to the sigmoidal kinetics observed with the intact holoenzyme. Moreover, the Scatchard plot for binding N-(phosphonacetyl)-L-aspartate (PALA) to the complex is linear with a Kd corresponding to that evaluated for the holoenzyme converted to the relaxed (R) state. Additional evidence that the binding of the zinc domain to the C trimer converts it to the R state was attained with a mutant form of ATCase in which Lys 164 in the catalytic chain is replaced by Glu. As shown previously (Newell, J.O. & Schachman, H.K., 1990, Biophys. Chem. 37, 183-196), this mutant holoenzyme, which exists in the R conformation even in the absence of active site ligands, has a 50-fold greater affinity for PALA than the free C subunit. Adding the zinc domain to the C trimer containing the Lys 164-->Glu substitution leads to a 50-fold enhancement in the affinity for the bisubstrate analog yielding a value of Kd equal to that for the holoenzyme. A different mutant ATCase containing the Gln 231 to Ile replacement was shown (Peterson, C.B., Burman, D.L., & Schachman, H.K., 1992, Biochemistry 31, 8508-8515) to be much less active as a holoenzyme than as the free C trimer. For this mutant holoenzyme, the addition of substrates does not cause its conversion to the R state. However, the addition of the zinc domain to the Gln 231-->Ile C trimer leads to a marked increase in enzyme activity, and PALA binding data indicate that the complex resembles the R state of the holoenzyme. This interaction leading to a more active conformation serves as a model of intergenic complementation in which peptide binding to a protein causes a conformational correction at a site remote from the interacting surfaces resulting in activation of the protein. This linkage was also demonstrated by difference spectroscopy using a chromophore covalently bound at the active site, which served as a spectral probe for a local conformational change. The binding of ligands at the active sites was shown also to lead to a strengthening of the interaction between the zinc domain and the C trimer.     

低碳发展背景下北京市土地利用与生态系统服务多情景模拟研究

在全球气候变化的背景下,合理协调低碳减排与经济效益之间的关系,进而指导土地利用优化配置,对于推动经济社会低碳转型与生态系统服务价值的提升具有重要意义。通过耦合多目标规划(MOP)与斑块生成土地利用模拟模型(PLUS),针对北京市构建了自然演变、经济优先与低碳发展这3种未来发展情景,并对不同情景下2030年土地利用与生态系统服务价值进行模拟评估。在此基础上,进一步对北京市未来生态系统服务提升区域进行识别,将传统的多情景土地利用模拟研究进行了延伸与扩展。结果表明：(1)3种情景下的土地利用结构与空间布局存在差异,低碳发展情景下东南部平原区建设用地的扩张与耕地面积的减少趋势得到了有效控制,西北部山区的林地面积有较大幅度的增长。(2)低碳发展情景下的生态系统服务价值总量约为590亿元,高于另外两种情景,且从空间分布上来看,该情景下服务价值提升区域的面积也显著高于其他情景。(3)低碳发展情景下,北京市生态系统服务提升潜力较大的区域面积占比约7.98%、潜力中等区域的面积占比约26.57%,潜力较低区域的面积占比约65.45%。     

Search for substrate-based inhibitors fitting the S2' space of malarial aspartic protease plasmepsin II.

Plasmepsin (Plm) has been identified as an important target for the development of new antimalarial drugs, since its inhibition leads to the starvation of Plasmodium falciparum. A series of substrate-based dipeptide-type Plm II inhibitors containing the hydroxymethylcarbonyl isostere as a transition-state mimic were synthesized. The general design principle was provision of a conformationally restrained hydroxyl group (corresponding to the set residue at the P2' position in native substrates) and a bulky unit to fit the S2' pocket.     

Identification of genes expressed in the shoot apex ofBrassica campestris during floral transition

The vegetative-to-floral transition ofBrassica campestris cv. Osome was induced by vernalization. Poly(A)+RNA was isolated from the transition shoot apex after 6 weeks of vernalization, the floral apex after 12 weeks of vernalization and the expanded leaves just before vernalization, and cDNAs were synthesized. These cDNAs were used for subtraction and differential screening to select cDNA preferentially present in the transition and floral apices. Nucleotide sequences of the resulting 14 cDNA clones were determined, and northern blot analysis was carried out on six cDNAs. Two cDNA clones which did not show significant similarity to known genes were shown to be preferentially expressed in the floral apex.     

Physicochemical basis for the rapid time-action of LysB28ProB29-insulin: dissociation of a protein-ligand complex.

The rate-limiting step for the absorption of insulin solutions after subcutaneous injection is considered to be the dissociation of self-associated hexamers to monomers. To accelerate this absorption process, insulin analogues have been designed that possess full biological activity and yet have greatly diminished tendencies to self-associate. Sedimentation velocity and static light scattering results show that the presence of zinc and phenolic ligands (m-cresol and/or phenol) cause one such insulin analogue, LysB28ProB29-human insulin (LysPro), to associate into a hexameric complex. Most importantly, this ligand-bound hexamer retains its rapid-acting pharmacokinetics and pharmacodynamics. The dissociation of the stabilized hexameric analogue has been studied in vitro using static light scattering as well as in vivo using a female pig pharmacodynamic model. Retention of rapid time-action is hypothesized to be due to altered subunit packing within the hexamer. Evidence for modified monomer-monomer interactions has been observed in the X-ray crystal structure of a zinc LysPro hexamer (Ciszak E et al., 1995, Structure 3:615-622). The solution state behavior of LysPro, reported here, has been interpreted with respect to the crystal structure results. In addition, the phenolic ligand binding differences between LysPro and insulin have been compared using isothermal titrating calorimetry and visible absorption spectroscopy of cobalt-containing hexamers. These studies establish that rapid-acting insulin analogues of this type can be stabilized in solution via the formation of hexamer complexes with altered dissociation properties.     

内蒙古草原退化群落恢复演替的研究II. 恢复演替时间进程的分析

 根据内蒙古典型草原地带的羊草+大针茅草原退化变型一冷蒿群落封育12年(1983—1994)的动态监测数据进行分析,对群落恢复演替轨迹取得以下认识： 1．依据群落优势种的更替及主分量分析结果可将恢复演替过程划分为冷蒿优势阶段、冷蒿+冰草阶段、冰草优势阶段、羊草优势阶段。 2;退化草原群落在恢复演替过程中,群落生产力的变化表现出阶梯式跃变和亚稳态阶面相间的特点。第一次跃变发生在1984年,上升到第二个阶面,第二次跃变发生在1990年,进入了第三个阶面,已接近于原生群落的生产力。 3．群落生产力与水资源量的关系因恢复演替阶段不同而异。第一亚稳态时期,群落地上现存生物量大体处于166g·m-2的水平上,生长季降水量达176mm以上时,增加降水对群落生产力的提高不发生显著影响。第二亚稳态时期,群落生物量与降水量之间的相关性显著。可推算出群落于物质生产用水量介于1.1～1.6mm·g-1之间。此值在1.1mm·g-1时,群落对水资源的利用效率最高,而在1.6mm·g-1时群落生物量达到最大值。 4．在恢复演替进程中,群落密度的位点常数约为271.5株·m-2,循此常数上下波动,表现出拥挤与稀疏交替发生的过程,构成了恢复演替的节奏性变化。群落生物量的跃变与亚稳态的形成,以及群落密度的拥挤与稀疏交替作用是群落恢复演替的内在机制。恢复演替的速度,到第10年发生了1.78个半变的生态距离。5．草原退化群落恢复演替过程中,按照其节奏性及生产力跃变与亚稳态的规律,调控放牧利用强度或采取技术措施,调节群落拥挤和稀疏的交替过程可加速恢复演替进程。     

The effect of income on the extraction of non-timber tropical forest products: Model,hypotheses, and preliminary findings from the Sumu Indians of Nicaragua

We use microeconomic theory to frame hypotheses about the effects of income on the use of non-timber rain forest products. We hypothesize that an increase in income: (a) encourages foraging specialization, resulting in the extraction of fewer goods; (b) increases the share of household income from occupations besides foraging; (c) produces a yearly value from the extraction of nontimber forest goods of about $50 per hectare; and (d) produces depletion of forest goods entering commercial channels and sustainable extraction of goods facing cheaper industrial substitutes. To examine these hypotheses we present worldwide ethnographic information and preliminary findings from field work carried out among the Sumu Indians of Nicaragua. Field work suggests that higher income produces: (a) foraging specialization with animals rather than with plants; (b) a decline in the economic importance of forest goods in household income; (c) and a rise in the value of non-timber goods removed from the forest to about $35/ha/year. We did not have time to test hypothesis d.     

Extracellular polysaccharide of Nostoc commune (Cyanobacteria) inhibits fusion of membrane vesicles during desiccation

Cells of the cyanobacterium Nostoc commune secrete a complex, high molecular weight, extracellular polysaccharide (EPS) which accumulates to more than 60% of the dry weight of colonies. The EPS was purified from the clonal isolate N. commune DRH1. The midpoint of the membrane phase transition (T_m) of desiccated cells of N. commune CHEN was low (T_{m dry} = 8 °C) and was comparable to the T_m of rehydrated cells((T_m)_H20 = 6 °C). The EPS was not responsible for the depression of T_m. However, the EPS, at low concentrations, inhibited specifically the fusion of phosphatidylcholine membrane vesicles when they were dried in vitro at0% relative humidity (−400 MPa). Low concentrations of a trehalose:sucrose mixture, in a molar ratio which corresponded with that present in cells in vivo, together with small amounts of the EPS, were efficient in preventing leakage of carboxyfloroscein (CF) from membrane vesicles. Freeze-fracture electron microscopy resolved complex changes in the structure of the EPS and the outer membrane in response to rehydration of desiccated cells. The capacity of the EPS to prevent membrane fusion, the maintenance of a low T_{m dry} in desiccated cells, and the changes in rheological properties of the EPS in response to water availability, constitute what are likely important mechanisms for desiccation tolerance in this cyanobacterium. This revised version was published online in August 2006 with corrections to the Cover Date.     

Plain tales from the grasslands: extraction, value and utilization of biomass in Royal BardiaNational Park, Nepal

Royal Bardia National Park in Southern Nepal contains a number of internationally endangered wildlife species, and a mix of habitats, including areas of grassland known as phantas. The conservation and current use of these areas is briefly described, including the extraction of grass from inside the National Park by local people living in adjacent villages. The products collected fulfil a range of human needs, both practical and ceremonial. The study attempts to quantify the annual offtake in both material and monetary terms, and comments on the sustainability of the practice. Alternative sources of roofing materials are apparently outside the scope of poorer households. The use of these materials has particular significance given the initiation of a Parks and People Project to implement a Buffer Zone around the National Park and effectively to link conservation and development needs.