具性别和阶段结构的非自治二种群捕食者-食饵系统研究

讨论了一类食饵具有性别结构,捕食者具有阶段结构的非自治捕食者．食饵系统,运用Liapunov函数方法,得到了该系统一致持续生存的充分条件．对于该模型的周期系统,在适当条件下,存在唯一、全局渐近稳定的周期解．对更具普遍意义的概周期现象,也得出了概周期正解唯一存在且全局渐近稳定的充分条件．     

The bioactive conformation of glucose-dependent insulinotropic polypeptide by NMR and CD spectroscopy

Glucose-dependent insulinotropic polypeptide (GIP) is a gastrointestinal incretin hormone, which modulates physiological insulin secretion. Because of its glucose-sensitive insulinotropic activity, there has been a considerable interest in utilizing the hormone as a potential treatment for type 2 diabetes. Structural parameters obtained from NMR spectroscopy combined with molecular modeling techniques play a vital role in the design of new therapeutic drugs. Therefore, to understand the structural requirements for the biological activity of GIP, the solution structure of GIP was investigated by circular dichroism (CD) followed by proton nuclear magnetic resonance (NMR) spectroscopy. CD studies showed an increase in the helical character of the peptide with increasing concentration of trifluoroethanol (TFE) up to 50%. Therefore, the solution structure of GIP in 50% TFE was determined. It was found that there was an alpha-helix between residues 6 and 29, which tends to extend further up to residue 36. The implications of the C-terminal extended helical segment in the inhibitory properties of GIP on gastric acid secretion are discussed. It is shown that the adoption by GIP of an alpha-helical secondary structure is a requirement for its biological activity. Knowledge of the solution structure of GIP will help in the understanding of how the peptide interacts with its receptor and aids in the design of new therapeutic agents useful for the treatment of diabetes.     

Solution structure and backbone dynamics of the AF-6 PDZ domain/Bcr peptide complex

The human AF-6, a scaffold protein between cell membrane-associated proteins and the actin cytoskeleton, plays an important role in special cell-cell junctions and signal transduction. It can be phosphorylated by the protein kinase Bcr, which allows efficient binding of the C terminus of Bcr to the PDZ domain of AF-6 and consequently enhances the binding affinity of AF-6 to Ras. Formation of the AF-6, Bcr, and Ras ternary complex results in down-regulation of the Ras-mediated signal transduction pathway. To better understand the molecular basis for the recognition of the AF-6 PDZ domain and Bcr, we solve the solution structure of the AF-6 PDZ domain complexed with the C-terminal peptide of Bcr and explore the interactions between them in detail. Compared with previously reported structures, the complex exhibits a noncanonical binding mode of PDZ/peptide. Owing to the distinct residues involved in the AF-6 PDZ domain and Bcr peptide interaction, the interaction mode does not adapt to the existing classification rules that have been put forward, based on the ligand or the PDZ domain specificity. Furthermore, the PDZ domain of AF-6 can bind to the C terminus of Bcr efficiently after phosphorylation of AF-6 by the Bcr kinase. The phosphorylation may induce a conformational change of AF-6, which makes the binding surface on the PDZ domain accessible to Bcr for efficient binding. This study not only characterizes the structural details of the AF-6 PDZ/Bcr peptide complex, but also provides a potential target for future drug design and disease therapy.     

Structural biology of transmembrane domains: efficient production and characterization of transmembrane peptides by NMR

Structural characterization of transmembrane peptides (TMPs) is justified because transmembrane domains of membrane proteins appear to often function independently of the rest of the protein. However, the challenge in obtaining milligrams of isotopically labeled TMPs to study these highly hydrophobic peptides by nuclear magnetic resonance (NMR) is significant. In the present work, a protocol is developed to produce, isotopically label, and purify TMPs in high yield as well as to initially characterize the TMPs with CD and both solution and solid-state NMR. Six TMPs from three integral membrane proteins, CorA, M2, and KdpF, were studied. CorA and KdpF are from Mycobacterium tuberculosis, while M2 is from influenza A virus. Several milligrams of each of these TMPs ranging from 25 to 89 residues were obtained per liter of M9 culture. The initial structural characterization results showed that these peptides were well folded in both detergent micelles and lipid bilayer preparations. The high yield, the simplicity of purification, and the convenient protocol represents a suitable approach for NMR studies and a starting point for characterizing the transmembrane domains of membrane proteins.     

2-Aminoethyl diphenylborinate analogues: selective inhibition for store-operated Ca2+ entry

Capacitative calcium entry (CCE), the mechanism that replenishes the internal Ca2+ stores with Ca2+ from the extracellular milieu in response to depletion of the store, is mediated by Ca2+ channels in the plasma membrane generally referred to as store-operated channels (SOCs). However, the roles of SOCs in the more physiological context have been fully elucidated. 2-Aminoethyl diphenylborinate (2-APB) strongly inhibits SOCs, as well as inositol-1,4,5 trisphosphate (IP3) receptors. In the present study, we screened a library of 166 2-APB analogues for effects on CCE and IP3-induced Ca2+ release in order to discover specific SOC inhibitors, and found that some blocked both store-operated and receptor-operated Ca2+ influx more strongly and selectively than 2-APB. Indeed, these new compounds ceased the prolonged intracellular Ca2+ oscillations induced by a low concentration of ATP in CHO-K1 cells. These novel SOC inhibitors will be valuable pharmacological and biochemical tools for elucidating the physiological roles.     

PEG模拟干旱胁迫对加拿大飞蓬种子萌发及幼根生长的影响

【目的】研究干旱胁迫对加拿大飞蓬种子萌发及幼根生长的影响,为科学防控加拿大飞蓬入侵提供依据。【方法】以当年生加拿大飞蓬成熟种子为材料,分别用蒸馏水（CK）和不同浓度5%、10%、15%、20%、25%的聚乙二醇溶液（PEG）（分别表示轻度、中度、重度、极度和极重度干旱胁迫强度）对加拿大飞蓬种子进行处理。测定日相对发芽率、发芽率、发芽势等指标,分析加拿大飞蓬种子萌发情况;测定幼苗根系宽、根系高、根平均直径、根面积、总根尖数及根分叉数,分析其幼苗根生长情况;并测定种子恢复萌发情况。【结果】无干旱胁迫的加拿大飞蓬种子萌发指标最高,随着干旱胁迫强度增加,种子萌发指标减小,极重度干旱胁迫下加拿大飞蓬种子不萌发。与CK相比,轻度干旱胁迫和重度干旱胁迫对加拿大飞蓬根平均直径影响不显著,但会促进侧根与不定根生长;重度干旱胁迫下,加拿大飞蓬的总根尖数和分叉数显著增加。加拿大飞蓬种子在PEG浓度为20.13%的干旱胁迫下不发芽。种子在极度和极重度干旱胁迫下不萌发,但当水分条件充足时,种子依然具有萌发的能力。【结论】建议在加拿大飞蓬种子尚未成熟前进行人工拔除,或在加拿大飞蓬入侵地种植耐旱强、生长快、根系发达、竞争性强的植物,以控制加拿大飞蓬的生长发育。     

Phase behavior of aqueous gelatin/oligosaccharide mixtures

The phase behavior of aqueous mixtures of gelatin and oligosaccharides above their gelation temperature is investigated experimentally, and rationalized according to a simple multicomponent Flory-Huggins model. When the gelatin is only weakly charged, entropic considerations dominate and it is found that the cloud point curve of the mixtures is extremely sensitive to the molecular weight distribution of the oligosaccharide. Even very small quantities of long-chain oligosaccharides present in an otherwise short-chain oligosaccharide population can radically reduce the compatibility. Added salt does not significantly affect the phase diagram, although a strong effect on the kinetics of phase separation is seen. Lowering the pH increases the electrostatic charge on the gelatin and strongly enhances the compatibility. Because the kinetics of gelation and phase separation are different, gelation can freeze in nonequilibrium states. Therefore, all phase diagrams were determined well above the gelation temperature (about 37°C). © 1997 John Wiley & Sons, Inc. Biopoly 41: 607–622, 1997     

Electrophoresis of proteins in semidilute polyethylene glycol solutions: Mechanism of retardation

The retardation of proteins in the M_t range of 15–500 kDa in capillary electrophoresis conducted in semidilute solutions of the polymer polyethylene glycol (M_t range 0.2–8.0 × 10⁶), was measured. The purpose was to test the predictions of the scaling theory with regard to the relation of retardation to (a) the M_t of the polymer, (b) the concentration of the polymer, and (c) the radius of the protein particles. These predictions derive from a mechanism that relates retardation to the screening length of the polymer solution, viewed as the average distance between the entanglement points of polymer chains. For the molecular weight range from 60 to 500 kDa of (near) spherical proteins, the retardation was found to be related to polymer concentration c asμ/μ_o = exp(-Ac^0.69)where μ/μ₀ is the retardation expressed as the ratio between the mobility in polymer solution and that in free solution. The value of the exponent of 0.69 is in close agreement with the value of 0.75 predicted by the scaling theory. Parameter A was found (a) to scale as the 0.04th power of M_t (polymer), approximating the predicted value of 0; and (b) to be proportional to particle radius as predicted. All measured values of retardation were independent of electric field strength in the range of 37–370 V/cm. Thus, experimental findings are consistent with the mechanism relating electrophoretic retardation to the screening length of the polymer network in the specified molecular weight range of proteins. Under the same conditions, log(μ/μ₀) of proteins with M_t's less than 60 kDa (a) scales as the −0.06th power of M_t (polymer), and (b) is proportional to polymer concentration, suggesting a retardation mechanism that is not related to the screening length. © 1997 John Wiley & Sons, Inc. Biopoly 42: 183–189, 1997     

Paramagnetic relaxation as a tool for solution structure determination: Clostridium pasteurianum ferredoxin as an example

The possibility of using the relaxation properties of nuclei for solution structure determination of paramagnetic metalloproteins is critically evaluated. First of all, it is theoretically and experimentally demonstrated that magnetization recovery in non-selective inversion recovery experiments can be approximated to an exponential in both diamagnetic and paramagnetic systems. This permits the estimate of the contribution of paramagnetic relaxation when dominant or sizable. Then, it is shown that the averaging of paramagnetic relaxation rates due to cross relaxation is often tolerably small with respect to the use of paramagnetic relaxation rates as constraints for structural determination. Finally, a protocol is proposed to use such paramagnetic relaxation rates, which depend on the sixth power of the metal to resonating nucleus distance, as constraints for solution structure determination of proteins. As an example, the available solution structure of the oxidized ferredoxin from Clostridium pasteurianum has been significantly improved in resolution especially in the proximity of the metal ions by using 69 new constraints based on paramagnetic relaxation. Proteins 29:348–358, 1997. © 1997 Wiley-Liss, Inc.