Intrachain disulfide bond in the core hinge region of human IgG4.

IgG is a tetrameric protein composed of two copies each of the light and heavy chains. The four-chain structure is maintained by strong noncovalent interactions between the amino-terminal half of pairs of heavy-light chains and between the carboxyl-terminal regions of the two heavy chains. In addition, interchain disulfide bonds link each heavy-light chain and also link the paired heavy chains. An engineered human IgG4 specific for human tumor necrosis factor-alpha (CDP571) is similar to human myeloma IgG4 in that it is secreted as both disulfide bonded tetramers (approximately 75% of the total amount of IgG) and as tetramers composed of nondisulfide bonded half-IgG4 (heavy chain disulfide bonded to light chain) molecules. However, when CDP571 was genetically engineered with a proline at residue 229 of the core hinge region rather than serine, CDP571 (S229P), or with an IgG1 rather than IgG4 hinge region, CDP571(gamma 1), only trace amounts of nondisulfide bonded half-IgG tetramers were observed. Trypsin digest reversephase HPLC peptide mapping studies of CDP571 and CDP571(gamma 1) with on-line electrospray ionization mass spectroscopy supplemented with Edman sequencing identified the chemical factor preventing inter-heavy chain disulfide bond formation between half-IgG molecules: the two cysteines in the IgG4 and IgG1 core hinge region (CPSCP and CPPCP, respectively) are capable of forming an intrachain disulfide bond. Conformational modeling studies on cyclic disulfide bonded CPSCP and CPPCP peptides yielded energy ranges for the low-energy conformations of 31-33 kcal/mol and 40-42 kcal/mol, respectively. In addition, higher torsion and angle bending energies were observed for the CPPCP peptide due to backbone constraints caused by the extra proline. These modeling results suggest a reason why a larger fraction of intrachain bonds are observed in IgG4 rather than IgG1 molecules: the serine in the core hinge region of IgG4 allows more hinge region flexibility than the proline of IgG1 and thus may permit formation of a stable intrachain disulfide bond more readily.     

Design, synthesis, expression, and characterization of the genes for mouse Fc gamma RIIb1 and Fc gamma RIIb2 cytoplasmic regions.

The cytoplasmic regions of the mouse low-affinity Fc gamma RII isoforms, mFc gamma RIIb1, and mFc gamma RIIb2, play a key role in signal transduction by mediating different cellular functions. mFc gamma RIIb1 has a 94-residue cytoplasmic region, whereas mFc gamma RIIb2 has a 47-residue cytoplasmic region. Genes encoding the cytoplasmic regions of mFc gamma RIIb1 (b1-94) and mFc gamma RIIb2 (b2-47) were designed, synthesized, and expressed as fusion proteins in Escherichia coli. A sequence-specific protease, thrombin, was used to release the b1-94 peptide, which was purified by using HPLC. The b2-47 peptide was synthesized chemically. CD spectropolarimetry was employed to examine the secondary structures of b1-94 and b2-47. These studies were conducted in aqueous solution, in mixtures of water and trifluoroethanol or methanol, and as a function of temperature. The results indicate that the b1-94 and b2-47 structures are sensitive functions of the solvent environment, and that nonaqueous solvents induce significant alpha-helical structure.     

Evolution of the mitochondrial DNA control region in the mbuna (Cichlidae) species flock of lake Malawi, East Africa

Considerable controversy has surrounded the application of mitochondrial DNA data to reconstruction of evolutionary relationships among the endemic cichlids of Lake Malawi. Central to this debate has been the issue of whether lineage sorting is complete, and thus whether these data actually reflect species phylogeny, or simply gene genealogy. Review of all mtDNA control region sequences available for members of one monophyletic subset of this species flock, the Malawi rockfishes, or mbuna, strongly indicates that lineage sorting is incomplete: Character-based analyses of these sequences reconstruct gene, not species, interrelationships. Analysis of the pattern of nucleotide substitutions differentiating these mtDNA alleles suggests that pyrimidine residues undergo transition substitutions more often than do purines. Estimation of the magnitude of derived sequence differentiation in light of the reconstructed gene genealogy suggests that the mbuna may be of considerably more recent vintage than previous molecular characterizations have indicated. Received: 6 April 1996 / Accepted: 3 March 1997     

The rDNA Internal Transcribed Spacer Region as a Taxonomic Marker for Nematodes

The ITS region from a wide taxonomic range of nematodes, including secernentean and adenophorean taxa, and free-living, entomopathogenic, and plant-parasitic species, was evaluated as a taxonomic marker. Size of the amplified product aided in the initial determination of group membership, and also suggested groups that may require taxonomic reevaluation. Congeneric species often displayed identically sized ITS regions, but genera such as Pratylenchus and Tylenchorhynchus had species with large differences in size. ITS heterogeneity in individuals and populations was identified in several nematode taxa. PCR-RFLP of ITS1 is advocated as a method of taxonomic analysis in genera such as Helicotylenchus that contain numerous species with few diagnostic morphological characteristics.     

Sex estimation using the first cervical vertebra

The articular surfaces and vertebral foramen area of the first cervical vertebra are sexually dimorphic and can be used to sex complete or fragmentary specimens. Eight measurements were taken from the articular regions (superior and inferior) of 100 first cervical vertebrae from Terry collection specimens housed at the Smithsonian Institution. Seven regression and seven discriminant function equations were created that predict sex with 77–85% and 75–85% accuracy, respectively. In separate control tests, measurements from 100 first cervical vertebrae from Hamann-Todd collection individuals (Cleveland Museum of Natural History) and from 34 archaeological specimens were used with the Terry equations. The control samples were sexed with 60—85% accuracy. © 1995 Wiley-Liss, Inc.     

毛茛科金莲花亚科植物的地理分布

本文对毛茛科金莲花亚科各属的地理分布作了分析,该亚科植物除了少数属的一些种分布到南半球的温带地区,一些种分布或延伸到亚热带山地、非洲东部和北部的干旱、半干旱的地区外,绝大部分的属、种均分布于泛北极区域。根据其17个属的地理分布式样,把它们划分为8个分布区类型:(1)北温带分布类型4属;(2)北温带和非洲分布类型1属;(3)北半球温带和南半球间断分布类型1属;(4)欧洲和东亚间断分布类型1属;(5)西亚分布类型1属;(6)地中海分布类型3属;(7)欧亚和温带亚洲分布类型1属;(8)东亚分布类型5属。本文以形态特征为主,结合花粉和染色体的性状分析,认为东亚特有的鸡爪草属、Megaleranthis和铁破锣属可能分别是联系驴蹄草属和金莲花属,鸡爪草属和金莲花属以及金莲花族和升麻族的中间类型。另外,文中详细地统计了该亚科的不同等级分类群及特有种在各个植物区的分布,并从系统发育的观点讨论了各个植物区所具有的原始类群和进化类群,提出了如下论点,即东亚植物区(特别是中国西南部)不但是金莲花亚科植物分布的多度和多样性中心以及特有类群的分布中心,而且还是原始类群的保存中心,伊朗-土兰区及地中海周围是第二分布中心。     

Mammalian expression of single chain variable region fragments dimerized by Fc regions

To promote application of a single chain variable region fragment (sFv) in immunoglobulins, a sFv gene was connected to an IgG1 Fc gene, designated as a sFvc gene, and used for transfection of Sp2/0. As a result, the sFvc protein was found to be secreted in a dimeric form. It is thus felt that the sFvc protein, which mimicks the shape of a naturally occurring antibody, can be simple and useful to reproduce divalency and Fc-associated effecter functions as seen in a natural antibody.Abbreviations Abbreviations sFv single chain variable region fragment - Fc constant region of immunoglobulin - sFvc single chain variable region fragment with an Fc region     

A conceptual model for the assessment of depressional wetlands in the prairie pothole region

The U.S. Environmental Protection Agency's (EPA's), Environmental Monitoring and Assessment Program (EMAP) is developing a landscape-level conceptual model to evaluate the condition of depressional (basin-type) wetlands in the prairie pothole region (PPR) of the United States. This effort is underway to determine the current condition of the Nation's wetlands and to track how it is improving or degrading over time, as well as to identify management priorities over major geographic areas. The depressional wetlands in the PPR were selected by EMAP both because of the importance of this region for waterfowl and because of the efforts currently being conducted by federal agencies and academic institutions in this region. The PPR provides nesting habitat for more than 15 species of ducks, and supports as much as half of the total production of dabbling and diving ducks in North America. Wetlands in this area became a vulnerable resource after extensive draineage in the 1800s. We propose a conceptual model that represents a framework for guiding the development of ecological indicators, research activities, and data collection for the evaluation of wetland conditions. In princple, this conceptual model is applicable to wetlands in any part of the world.