A new approach for rhizosphere research by X-ray microanalysis of microliter soil solutions

Lolium perenne growing with high root density on a fine nylon mesh (Kuchenbuch and Jungk, 1982) caused the development of element gradients in the rhizosphere below the mesh. Micro-liter soil solutions from 2-mg soil samples were sprayed onto Formvar-coated grids and analyzed by X-ray microanalysis in a transmission electron microscope. The results were comparable to those obtained by flame photometry and atomic absorption spectrometry (AAS) of conventional soil solutions from 1 g soil. X-ray microanalysis of micro-soil solutions allows the application of different extraction procedures to even small amounts of soil usually available from rhizosphere experiments. Information about soil buffering characteristics in the rhizosphere can thus be obtained. Aluminum accumulation in the rhizosphere of small segments of single Picea abies fine roots grown in undisturbed natural forest soil could be detected with this technique.     

The effects of continuous illumination on southern and northern Micrasterias strains

Different strains of Micrasterias (Chlorophyta, Conjugatophyceae); M. rotata (Grev.) Ralfs ex. Ralfs and M. denticulata Breb. ex. Ralfs var. angulosa (Hantzsch) W. & G. S. West from northern and southern Finland were treated with continuous illumination in order to study the cellular effects of the treatment and whether the tolerance to continuous light of the northern Finnish strains is related to the different daylenght conditions in northern and southern areas. During the growing season the Finnish strains normally live in long-day conditions or even in continuous light (between 60 and 70°N), and they also tolerated continuous illumination in the laboratory. Ultrastructural changes were found especially in the chloroplasts, where formation of calcium precipitates of different forms and sizes and also formation of plastoglobuli containing lipids appeared. However, even in 4-week treatments the ultrastructure of cells of these northern strains was not totally disrupted, contrary to what was found in southern M. torreyi , studied earlier. Southern and northern strains tolerated continuous illumination in different ways. They seem to differ from each other physiologically, and the differences are possibly located in their ionic metabolism and regulation. The injuries sustained during continuous illumination of Micrasterias may largely be caused by the accumulation of Ca²⁺ in cytoplasm and organelles, especially in the chloroplasts.     

Structural studies of reagents for peptide bond formation: Crystal and molecular structures of HBTU and HATU

Summary X-ray structure determinations of HBTU and HATU, well-known reagents for peptide bond formation, show that the solid-state structures of these compounds differ markedly from those commonly presented in the literature. The solid-state structures are isomers of the N,N,N',N'-tetramethyluronium salt formulation commonly written for HBTU and HATU. HBTU and HATU, obtained either directly from synthesis or from CH₃CN solution, crystallize as the guanidinium N-oxide isomers. Both compounds crystallize in nearly identical conformations, despite marked differences in crystal packing.     

Peptide crystal growth via sulfonate salt formation: The structure of bis-glycylglycine-1,5-naphthalenedisulfonate hydrate

Summary Continuing a line of investigations on methods for formation and growth of high-quality crystals of peptides, the glycylglycine sequence has been crystallized by evaporation methods as a salt with 1,5-naphthalenedisulfonic acid. The structure of the peptide is highly extended, and is conformationally quite similar to the structures which have been characterized for other zwitterionic and salt forms of this sequence. Thus, crystallization as a salt with this sulfonic acid has imposed no undue influence upon the molecular conformation. These results offer further indication that the preparation of peptide sulfonate salts, particularly with arene templates, may have broad general utility for crystallization of interesting sequences which until now have not been approachable in their zwitterionic forms.     

1.56 Å structure of mature truncated human fibroblast collagenase

The X-ray crystal structure of a 19 kDa active fragment of human fibroblast collagenase has been determined by the multiple isomorphous replacement method and refined at 1.56 Å resolution to an R-factor of 17.4%. The current structure includes a bound hydroxamate inhibitor, 88 waters and three metal atoms (two zincs and a calcium). The overall topology of the enzyme, comprised of a five stranded β-sheet and three α-helices, is similar to the thermolysin-like metalloproteinases. There are some important differences between the collagenase and thermolysin families of enzymes. The active site zinc ligands are all histidines (His-218, His-222, and His-228). The presence of a second zinc ion in a structural role is a unique feature of the matrix metalloproteinases. The binding properties of the active site cleft are more dependent on the main chain conformation of the enzyme (and substrate) compared with thermolysin. A mechanism of action for peptide cleavage similar to that of thermolysin is proposed for fibroblast collagenase. © 1994 Wiley-Liss, Inc.     

A surface mutant (G82R) of a human α-glutathione S-transferase shows decreased thermal stability and a new mode of molecular association in the crystal

A chimeric enzyme (GST121) of the human α-glutathione S-transferases GST1-1 and GST2-2, which has improved catalytic efficiency and thermostability from its wild-type parent proteins, has been crystallized in a space group that is isomorphous with that reported for crystals of GST1-1. However, a single-site (G82R) mutant of GST121, which exhibits a significant reduction both in vitro and in vivo in protein thermostability, forms crystals that are not isomorphous with GST1-1. The mutant protein crystallizes in space group P2₁2₁2₁, with cell dimensions a = 49.5, b = 92.9, c = 115.9 Å, and one dimer per asymmetric unit. Preliminary crystallographic results show that a mutation of the surface residue Gly 82 from a neutral to a charged residue causes new salt bridges to be formed among the GST dimers, suggesting that the G82R mutant might aggregate more readily than does GST121 in solution resulting in a change of its solution properties. © 1994 Wiley-Liss, Inc.     

Zn对细胞保护作用机理的研究

应用扫描质子微探针和同步辐射ｘ荧光分析技术测定了细胞中元素的分布和组成,为确定Zn是细胞结构成分提供了直接的实验依据.用上述核技术结合有关生化指标,分析测定了正常和损伤细胞（脂质过氧化损伤）中Fe,Zn和丙二醛、SH基含量变化的相互关系.实验结果表明,当细胞发生脂质过氧化损伤时,Fe含量和丙二醛含量同步增高,而Zn含量和SH基量则降低.给细胞补充Zn后,提高了细胞质膜中的Zn含量,SH基量也随之增加,同时丙二醛量降低.提示Zn保护细胞完整性的作用机理之一是控制脂质过氧化作用.Zn可保护膜蛋白的SH基,减少和阻止被Fe所催化的过氧化反应.     

The structure of duplex DNA in the nucleosome core particle: An alternative model

Many studies indirectly indicate that the conformation ofin vivo duplex DNA is the double helix. The most direct view, from the X-ray analysis of the nucleosome core particle, has also been interpreted in terms of the double helix structure. However, an alternative possibility exists; that the duplex adopts a metastable side-by-side conformation which readily converts to the double helix on removal of protein. Evidence for the existence of this conformation has been obtained from a reanalysis of the electron density map for the nucleosome particle.     

乌骨鸡黑素的一些基本结构特征的初步研究

应用元素分析、电子能谱(XPS)和红外光谱(IR)分析技术对乌骨鸡体内黑素结构特征进行了研究。元素分析结果得到黑素分子中C、H、N、O克原子比率的均值为10:12:1:3.该比值与动物性吲哚黑素的相应元素比例接近。XPS波谱显示出黑素中含有两种价态的硫(S~Ⅵ、S~Ⅰ),其中以低价态占优势,S2p电子结合能比人发黑素要大些。氮原子只有一种价态。IR光谱中有五组明显吸收峰,其中以主要表征吲哚环的1619—1632CM~(-1)处的吸引峰最强。以上结果初步表明出乌骨鸡黑素是以吲哚环为主体的异聚物。