The role of tyrosine-114 in the enzymatic activity of the Shiga-like toxin I A-chain

Shiga-like toxin I (SLT-I), the potent cytotoxin produced by certain pathogenic strains of Escherichia coli, is a member of a burgeoning family of ribosome-inactivating proteins (RIPS), which share common structural and mechanistic features. The prototype of the group is the plant toxin ricin. Recently we proposed a structural model for the Slt-IA active site, based in part on the known geometry of the enzymatic subunit of the ricin toxin. The model places three aromatic residues within the putative Slt-IA active site cleft: tyrosine 77, tyrosine 114, and tryptophan 203. Here we present biochemical and biophysical data regarding, the phenotypes of conservative point mutants of Slt-IA in which tyrosine 114 is altered. We used oligonucleotide-directed mutagenesis to replace tyrosine 114 with either phenylalanine (Y114F) or serine (Y114S). Periplasmic extracts of E. coli containing wild-type or mutant Slt-IA were tested for their ability to inhibit protein synthesis in vitro. Relative to wild-type, the activity of mutant Y1 14F was attenuated about 30-fold, while the mutant Y114S was attenuated about 500 to 1000-fold. In order to address the possibility that differential activation of the mutants rather than local effects at the active site might account for their diminished activity, we engineered the same mutations into a truncated slt-IA cassette that directs expression of a product corresponding to the activated A₁ form of Slt-IA (wild-type-). The same general relationships held: relative to wild type-, Y114F- was attenuated about 7-fold, and Y114S- about 300-fold. Tryptic digestion profiles of the mutant proteins were similar to those of the corresponding wild-type, indicating that the amino acid substitutions had not caused major alterations in conformation. We conclude that Y114 plays a significant role in the activity of Slt-IA, one which is quantitatively similar to that of Y77, and one which is predicated on the presence of both its weakly acidic phenolic hydroxyl and its aromatic ring.     

Gastrin and cholecystokinin in the Eastern Grey kangaroo, Macropus giganteus giganteus

Gastrin and CCK are believed to have a common ancestor. The gastrin structure has probably evolved from CCK-like peptides at a stage later than the amphibians. To trace the evolution of gastrin and CCK we have determined their structures in an Australian marsupial, the Eastern Grey kangaroo. The brain CCK was identical to CCK-8 of most mammals. The larger form of kangaroo gastrin was a 33mer with the sequence pELHPQDLPHLMTDLSKKKGPWQEEDAAY(SO₃)GWMDF-NH₂. The 11 italic residues indicate differences from human gastrin. Gastrin-15 and gastrin-16 comprised about 70% of the total immunoreactivity and resulted from cleavage after the second and third residue, respectively, of the unusual tribasic cleavage site. The smaller forms of kangaroo gastrin differ from most other mammalian gastrins in that the N-termini are not blocked with a pyroGlu moiety. Unlike other gastrins, kangaroo gastrin is more than 95% sulfated. The present study indicates that the gastrin structure, as defined by having a Tyr at position 6 from the C-terminus, evolved before the marsupials diverged from the other mammals 130 million years ago.     

不列颠·哥伦比亚省北部小杆松及白云杉立地指数与生态立地质量的关系

为在ＰｒｉｎｃｅＲｕｐｅｒｔ林区的“亚北方”部分建立生态立地质量与森林生产力的联系,对从９３个小杆松林分和７７个白云杉林分获得的数据进行了分析．所研究的林分处于两个气候状况、８个土壤水分状况以及５个土壤养分状况．这些气候、土壤水分和养分状况被视为等级变量用于林地分类和回归分析．小杆松和白云杉的立地指数随土壤水分和养分状况变化而变化,但不依赖于气候变化．与土壤水分相关的变化格局对两个种来说很相似,但与土壤养分相关的变化格局则全然不同．在所建立的５类回归模型中,土壤小区模型对于两个种都显示出立地指数与土壤水分和养分状况具有很强的相互关系（Ｒ２＞０．８０,ＳＥＥ≤１．６ｍ）．可以认为土壤水分和养分的等级度量在大范围内可作为小杆松和白云杉立地指数的预测预报因子．     

The SKS of the KMSKS signature of class I aminoacyl-tRNA synthetases corresponds to the GKT/S sequence characteristic of the ATP-binding site of many proteins

On the basis of protein modification studies and primary structure comparison, we propose that the SKS sequence within the KMSKS signature of the class 1 aminoacyl-tRNA synthetases corresponds to the GKT(or S) sequence considered as a signature of the nucleotide triphosphate-binding site of many proteins.     

Site fidelity in predictable and unpredictable habitats

Summary Site fidelity, the tendency to return to a previously occupied location, has been observed in numerous species belonging to at least three phyla. In this paper I develop a general model using dynamic programming to investigate conditions under which fidelity to a previously occupied territory will be advantageous. The results predict that site fidelity should be inversely related to heterogeneity in territory quality and the animal's lifespan and positively related to the cost of changing territories, age and probability of mortality in the habitat. The predictability of reproductive outcome (defined as the probability that next period's outcome will be the same as this period's outcome) also affects site fidelity. In predictable habitats, changing territories may be favoured after a bad previous outcome. In contrast, settlement should be independent of the previous outcome in unpredictable habitats. Individuals should also be site-faithful in unpredictable habitats, as long as the mean territory quality is equal among available territories. I also investigate the success of two potential decision rules (always stay and win-stay: lose-switch) relative to the optimal settlement strategy. The results show that these rules may perform as well as the optimal strategy under certain conditions. The always stay strategy does well in unpredictable habitats, when the mean quality within a territory is equal among territories. In contrast, the win-stay: lose-switch strategy performs best in predictable habitats.     

Defining null expectations for animal site fidelity

Site fidelity—the tendency to return to previously visited locations—is widespread across taxa. Returns may be driven by several mechanisms, including memory, habitat selection, or chance; however, pattern-based definitions group different generating mechanisms under the same label of ‘site fidelity’, often assuming memory as the main driver. We propose an operational definition of site fidelity as patterns of return that deviate from a null expectation derived from a memory-free movement model. First, using agent-based simulations, we show that without memory, intrinsic movement characteristics and extrinsic landscape characteristics are key determinants of return patterns and that even random movements may generate substantial probabilities of return. Second, we illustrate how to implement our framework empirically to establish ecologically meaningful, system-specific null expectations for site fidelity. Our approach provides a conceptual and operational framework to test hypotheses on site fidelity across systems and scales.     

Restudy of the upper age limit of Beijing Man site

The chronological position of Beijing Man site was restudied by U-series dating of two calcite samples taken from a stalagmitic flowstone layer intercalated in 1–2 layers. Based on the weighted mean of three measurements performed on the purer one, the upper age limit of this site be 421 ₋₅₄ ⁺¹¹⁰ ka B.P., much greater than former evaluation of 230 ka based on U-series dating of fossil bones. The reliability of the obtained results was discussed.     

Species of Borrelia distinguished by restriction site polymorphisms in 16S rRNA genes

Abstract Three phyletic groups of Borrelia associated with Lyme disease, B. burgdorferi, B. garinii and group VS461 can be distinguished from each other and other species of Borrelia by Bfa I restriction site polymorphisms in PCR amplified 16S rRNA genes. One strain isolated from an Ixodes pacificus tick in California that was previously unclassifiable was distinguishable from B. burgdorferi by an Mnl I restriction site polymorphism.     

低温与光对黄瓜幼苗子叶光合电子传递活性的影响

25～30℃和30 μmol m~(-2)s~(-1)光下培养的黄瓜幼苗,在黑暗下经 1～7℃处理24h或5℃处理24～72h,光合电子传递活性受不同程度的抑制;其抑制部位主要在PSⅡ氧化侧;随温度的降低和时间的延长,抑制部位可发展至PSⅡ及之后的电子递体上,但尚未影响PSⅠ的活性。160μmol m~(-2)s~(-1)的光强加重低温对电子传递活性的抑制,光强越高,则加重的程度越高;抑制部位从PSⅡ氧化侧发展至PSⅡ反应中心以及PSⅠ。