Nuclear Magnetic Resonance Studies on Huwentoxin-XI from the Chinese Bird Spider Ornithoctonus huwena： ^15N Labeling and Sequence-specific ^1H, ^15N Nuclear Magnetic Resonance Assignments

Huwentoxin-XI purified from the Chinese bird spider Ornithoctonus huwena is a toxin with both antiprotease activity and potassium channel blocking activity. To determine its solution structure, huwentoxin-XI was expressed in a yeast eukaryotic expression system and studied by NMR. The ^15N labeling strategy was used to facilitate the process of resonance assignments. The nearly complete sequence-specific assignments of proton and nitrogen resonances were obtained by analyzing a series of two-dimensional （2D） and three-dimensional （3D） spectra, including DQF-COSY, TOCSY, NOESY, ^15N-^1H HSQC, ^15N-^1H HNHA, ^15N-^1H HNHB, ^15N-^1H TOCSY-HSQC and ^15N-^1H NOESY-HSQC spectra. Secondary structure analysis of huwentoxin-XI showed that it mainly contains an N-terminal 310-helix from Thr3 to Arg5 and a C-terminal α-helix from Gln45 to Cys52, plus a triple-stranded antiparallel β-sheet of Glu18-Asn23, Thr26-Ile31 and Asn40-Lys41. These studies provide a solid basis for the final structure determination of huwentoxin-XI.     

Studies on the Chinese Woodsiaceae (3)—Phylogeny and Speciation

The history of the study on the woodsiodes is briefly surveyed in the paper. The family Woodsiaceae is recognized by the present author. The relationships among the species and the probable evolution of the family are discussed based on author's cytological and comparative morphological studies, and are indicated by Wagner's method, with numerical values as the indices. Woodsiaceae may have originated from the common ancestor of modern Dicranopteris and Sticherus of Gleicheniaceae, and evolved from it into two main branches, i.e., Woodsia and Protowoodsia. The origin of species through polyploid series are discussed, and W. andersonii, W. subcordata, W. alpina and Cheilanthopsis indosiosa considered as fertile allopolyploids; the probable way of speciation is also suggested for these species.     

Letter to the Editor: Backbone 1H, 15N and 13C Assignments for the Subunit a of the E. Coli ATP Synthase

The structure of the 30 KDa subunit a of the membrane component (F(0)) of E. coli ATP synthase is investigated in a mixture of chloroform, methanol and water, a solvent previously used for solving the structure of another integral membrane protein, subunit c. Near complete backbone chemical shift assignments were made from a set of TROSY experiments including HNCO, HNCA, HN(CA)CB, HN(CO)CACB and 4D HNCOCA and HNCACO. Secondary structure of subunit a was predicted from the backbone chemical shifts using TALOS program. The protein was found to consist of multiple elongated alpha-helical segments. This finding is generally consistent with previous predictions of multiple transmembrane alpha-helices in this polytopic protein.     

Validation of Drug-Like Inhibitors against Mycobacterium Tuberculosis L-Aspartate α-Decarboxylase Using Nuclear Magnetic Resonance (1H NMR)

The catalytic activity of L-aspartate α-decarboxylase (ADC) is essential for the growth of several micro-organisms, including Mycobacterium tuberculosis (Mtb), and has triggered efforts for the development of pharmaceutically active compounds against tuberculosis. The present study is a continuation of our recent chemoinformatics-based design approach for identifying potential drug-like inhibitors against MtbADC. We report an NMR-based protocol that allows label-free and direct monitoring of enzymatic conversion, which we have combined with a systematic testing of reported and newly identified potential inhibitors against MtbADC. Quantification of enzymatic conversion in the absence and presence of inhibitors allowed for a relative measure of the inhibitory effect (k_rel). Among the newly identified compounds, D-tartrate, L-tartrate, and 2,4-dihydroxypyrimidine-5-carboxylate were found to inhibit the enzyme with k_rel values of 0.36, 0.38, and 0.54, respectively. In addition to the identification of potential building blocks for the development of therapeutic agents, the current study highlights the importance of electrostatic interactions governing enzyme-inhibitor binding.     

Systematic Studies on Chinese Stachyuraceae (1)—Phytogeographical,Cytological, Palynological

The present paper, mainly dealing with phytogeographical, Cytological and Palynolo gical studies on Chinese Stachyuraceae, is the first part of a series of contributions to systematics of this family. As we know, Stachyuraceae, a monogeneric family, is endemic to East Asia, with a strong differentiation in China. Systematic position of the family is still uncertain. Undoubtedly, multidisciplinary studies on it wisl give a better understanding of plant differentiation in East-Asiatic floristic region and systematic position of the family. The distribution of all the 9 Chinese species of Stachyurus have been studied on the basis of morphogeographical method. The status of some taxa has been changed as follows: Stachyurus chinensis Franch. ssp. latus (Li) Y. C. Tang et Y. L. Cao, stat. nov. S. chinen-sis Franch. Ssp. cuspidatus (Li) Y. C. Tang et Y. L. Cao, stat. nov. S. chinensis Franch. ssp. brachystachyus (C. Y. Wu et S. K. Chen) Y. C. Tang et Y. L. Cao, stat. nov. S. salicifolius Franch. ssp. lancifolius (C. Y. Wu) Y. C. Tang et Y. L. Cao, stat. nov., S. yunnanensis Franch. var. pedicellatus Rehd. and S. oblongifolius Wang et Tang being reduced as synonyms of S. yunnanensis Franch. From morphological point of view, we would consider that sect. Callosurus is more primitive and its present area might be the preservation center of the family. In this paper the chromosome numbers of 5 taxa, i.e. Stachyurus yunnanensis, S. chinensis ssp. chinensis, S. chinensis ssp. cuspidatus, S. salicifolius ssp. salicifolius, S. retusus, are reported for the first time. Along with the chromosome countings of S. himalaicus and S. sigeyosii by S. kurosawa, we come to the conclusion that all the species so far studied have 24 small somatic chromosomes (2n=24). The pollen grains of 6 taxa, i.e. Stachyurus yunnanensis, S. obovatus, S. himalaicus, S. chinensis ssp. chinensis, S. salicifolius ssp. salicifolius, S. retusus were also examined. Together with the observations of G. Erdman and Huang Tsengchien on S. himalaicus, it seems that the pollen grains of the genus Stachyurus are rather uniform in morphology. The pollen grains of sect. Callosurus are tricolporoidate, while those of sect. Stachyurusare tricolporate. Two pollen types, representeb by the two sections, though not strongly distinct, may be considered as further evidence of the validity of the two taxa as section. Sexuality of flowers is also disscussed in the paper. According to our observationon six species, i.e. Stachyurus yunnanensis, S. obovatus, S. himalaicus, S. chinensis, S. salicifolius, S. retusus, the plants are dioecious, which supports T. Makino's observation on S. praecox and H. Hara's on S. himalaicus. Female flowers have relatively large pistils and short stamens with empty anthers, while male flowers have smaller sterile pistils and longer stamens with well developed anthers. Decontaminated thianthrene disproportion. 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Sequence-specific backbone 1H, 13C and 15N assignments of the 34 kDa catalytic domain of PTPN5 (STEP)

PTPN5 is a protein tyrosine phosphatase that plays an integral role in regulating excitatory postsynaptic activity. The sequence-specific backbone assignments of the murine PTPN5 catalytic domain have been determined based on triple-resonance experiments using uniformly [²H,¹³C,¹⁵N]-labeled protein.     

Assignments of backbone 1H, 13C and 15N resonances in H-Ras (1–166) complexed with GppNHp at physiological pH

The small GTPase Ras is an important signaling molecule acting as a molecular switch in eukaryotic cells. Recent findings of global conformational exchange and a putative allosteric binding site in the G domain of Ras opened an avenue to understanding novel aspects of Ras function. To facilitate detailed NMR studies of Ras in physiological solution conditions, we performed backbone resonance assignments of Ras bound to slowly hydrolysable GTP mimic, guanosine 5′-[ß, γ-imido]triphosphate at pH 7.2. Out of 163 non-proline residues of the G domain, signals from backbone amide proton, nitrogen and carbon spins of 127 residues were confidently assigned with the remaining unassigned residues mostly located at the exchange-broadened effectors interface.     

Patterns of δ~(13) C and δ~(15) N in wolverine Gulo gulo tissues from the Brooks Range,Alaska

Knowledge of carnivore diets is essential to understand how carnivore populations respond demographically to variations in prey abundance.Analysis of stable isotopes is a useful complement to traditional methods of analyzing carnivore diets.We used data on δ 13 C and δ 15 N in wolverine tissues to investigate patterns of seasonal and annual diet variation in a wolverine Gulo gulo population in the western Brooks Range,Alaska,USA.The stable isotope ratios in wolverine tissues generally reflected that of terr...     

Sequence-specific 1H, 13C and 15N backbone resonance assignments of the 34 kDa catalytic domain of human PTPN7

PTPN7 is a protein tyrosine phosphatase responsible for inactivation of MAPK in leukocytes. Here we report the backbone resonance assignments of the 34 kDa phosphatase domain of human PTPN7, which is amplified in myeloid malignancies and deleted in lymphoproliferative disorders.