The enzyme lactate dehydrogenase as a structural protein in avian and crocodilian lenses

The major components of mammalian lenses are tissue-specific, soluble proteins, the alpha-, beta- and gamma-crystallins. The lenses of other vertebrate classes often contain other major proteins, notably delta-crystallin in birds and reptiles. A fourth distinct type, described as epsilon-crystallin, is prominent in many bird and crocodile lenses. Here we show that epsilon-crystallin is an active glycolytic enzyme, lactate dehydrogenase (LDH) (EC 1.1.1.27) and that duck epsilon-crystallin appears to be identical to duck LDH-B4. LDH is a normal metabolic component in other lenses, but in duck is present in amounts far exceeding the requirements of any likely catalytic role. It appears that an active enzyme has been recruited, unchanged, to an extra role as a structural protein in the lens without gene duplication and sequence divergence. This surprising discovery raises the possibility that other crystallins may similarly be enzymes expressed at high levels in lens as structural proteins.     

Hedgehog signalling controls eye degeneration in blind cavefish

Hedgehog (Hh) proteins are responsible for critical signalling events during development but their evolutionary roles remain to be determined. Here we show that hh gene expression at the embryonic midline controls eye degeneration in blind cavefish. We use the teleost Astyanax mexicanus, a single species with an eyed surface-dwelling form (surface fish) and many blind cave forms (cavefish), to study the evolution of eye degeneration. Small eye primordia are formed during cavefish embryogenesis, which later arrest in development, degenerate and sink into the orbits. Eye degeneration is caused by apoptosis of the embryonic lens, and transplanting a surface fish embryonic lens into a cavefish optic cup can restore a complete eye. Here we show that sonic hedgehog (shh) and tiggy-winkle hedgehog (twhh) gene expression is expanded along the anterior embryonic midline in several different cavefish populations. The expansion of hh signalling results in hyperactivation of downstream genes, lens apoptosis and arrested eye growth and development. These features can be mimicked in surface fish by twhh and/or shh overexpression, supporting the role of hh signalling in the evolution of cavefish eye regression.     

Maintenance of optical quality during crystalline lens growth

Vertebrate lenses grow throughout life by the division of cells at the lens surface. The fibre cells thus produced are gradually covered by newer tissue, giving a layered structure. During growth, the lens must remain transparent and retain its refractile properties. The severity of these constraints is perhaps most evident in teleost fish which have a spherical lens that may increase in volume by a thousandfold during the first year of life. The dioptric power of the teleost fish eye is vested entirely in this spherical lens, as water, the cornea and the intraocular vitreous humour have almost identical refractive indices. Spherical lenses of uniform refractive index produce poor images because rays entering at different distances from the optic axis are focused at different distances from the lens. Teleost fish do not suffer from this imperfection and it has long been presumed that this is because there exists a refractive index gradient having a high value in the centre and decreasing continuously and symmetrically with radius in all directions. Here we demonstrate in the African cichlid fish, Haplochromis burtoni, that a refractive index gradient does exist, although its form is significantly different from that previously postulated.     

障复明滴眼液对半乳糖性白内障的防治作用

用半乳糖诱发大鼠白内障,研究障复明滴眼液对白内障的防治作用．测定了晶体内的可溶性蛋白,ＳＯＤ（超氧化物歧化酶）,ＧＳＨ（谷胱甘肽）和ＭＤＡ（丙二醛）水平．结果表明,障复明滴眼液具有保护晶体抗氧化防御体系的功能,能延缓和控制白内障的形成,对早期白内障有一定的治疗作用．     

A new method of research on molecular evolution of proteinase superfamily

The molecular evolutionary tree, also known as a phylogenetic tree, of the serine proteinase superfamily was constructed by means of structural alignment. Three-dimensional structures of proteins were aligned by the SSAP program of Orengo and Taylor to obtain evolutionary distances. The resulting evolutionary tree provides a topology graph that can reflect the evolution of structure and function of homology proteinase. Moreover, study on evolution of the serine proteinase superfamily can lead to better understanding of the relationship and evolutionary difference among proteins of the superfamily, and is of significance to protein engineering, molecular design and protein structure prediction. Structure alignment is one of the useful methods of research on molecular evolution of protein.     

Protein dispensability and rate of evolution.

If protein evolution is due in large part to slightly deleterious amino acid substitutions, then the rate of evolution should be greater in proteins that contribute less to individual fitness. The rationale for this prediction is that relatively dispensable proteins should be subject to weaker purifying selection, and should therefore accumulate mildly deleterious substitutions more rapidly. Although this argument was presented over twenty years ago, and is fundamental to many applications of evolutionary theory, the prediction has proved difficult to confirm. In fact, a recent study showed that essential mouse genes do not evolve more slowly than non-essential ones. Thus, although a variety of factors influencing the rate of protein evolution have been supported by extensive sequence analysis, the relationship between protein dispensability and evolutionary rate has remained unconfirmed. Here we use the results from a highly parallel growth assay of single gene deletions in yeast to assess protein dispensability, which we relate to evolutionary rate estimates that are based on comparisons of sequences drawn from twenty-one fully annotated genomes. Our analysis reveals a highly significant relationship between protein dispensability and evolutionary rate, and explains why this relationship is not detectable by categorical comparison of essential versus non-essential proteins. The relationship is highly conserved, so that protein dispensability in yeast is also predictive of evolutionary rate in a nematode worm.     

Ancestral lysozymes reconstructed, neutrality tested, and thermostability linked to hydrocarbon packing

The controversy surrounding the idea that neutral mutations dominate protein evolution is attributable in part to the inadequacy of the tools available to evolutionary investigators. With a few exceptions, most investigations into the force driving protein evolution have relied on indirect criteria for distinguishing neutral and non-neutral variants. To investigate a particular pathway of molecular evolution, we have reconstructed by site-directed mutagenesis likely ancestral variants of the lysozymes of modern game birds (order Galliformes), tested their activity and thermostability and determined their three-dimensional structure. We focused on amino acids at three positions that are occupied in all known game birds either by the triplet Thr 40, Ile 55, Ser 91, or by the triplet Ser 40, Val 55, Thr 91. We have synthesized proteins representing intermediates along the possible three-step evolutionary pathways between these triplets. Although all of these are active and stable, none of these intermediates is found in known lysozymes. A comparison of the structures and thermostabilities of the variants reveals a linear correlation between the side-chain volume of the triplet and the thermostability of the protein. Each pathway connecting the two extant triplet sequences includes a variant with a thermostability outside the range of the extant proteins. This observation is consistent with a non-neutral evolutionary pathway. The existence of variants that are more stable than the extant proteins suggests that selection for maximum thermostability may not have been an important factor in the evolution of this enzyme.     

Myxococcus xanthus spore coat protein S may have a similar structure to vertebrate lens beta gamma-crystallins

The Gram-negative bacterium Myxococcus xanthus has a complex life cycle during which large amounts of a protein of relative molecular mass (Mr) 19,000, known as protein S, are assembled into a spore surface coat by a process that specifically requires calcium ions. The gene for protein S has been cloned and the DNA sequence shows that the gene product is composed of four internally repeated homologous sequences, each 40 amino acids long. Although protein S resembles calmodulin both in its internally duplicated structure and its ability to bind calcium, it apparently has a beta-sheet secondary structure rather than the helix-loop-helix motifs that characterize the calmodulin family. We now show that protein S has a striking homology with the beta- and gamma-crystallins of the vertebrate eye lens which are beta-sheet proteins with internally duplicated structures. This implies that the beta- and gamma-crystallins evolved from already existing proteins, whose ancestors occurred in the prokaryotes. The biological function of protein S, as a closely packed, stable protein in a relatively dehydrated environment, has implications for the functions of crystallins, which are found closely packed in the lens fibre cells, where their stability is essential for maintenance of transparency.     

谷胱甘肽硫转移酶结构与功能研究进展

谷胱甘肽转硫酶(G lutath ione S-transferases,简称GSTs,EC2.5.1.18)是广泛分布于哺乳动物、植物、鸟类、昆虫、寄生虫及微生物体内的一组多功能同工酶,其主要功能是催化某些内源性或外来有害物质的亲电子基团与还原型谷胱甘肽的巯基偶联,增加其疏水性使其易于穿越细胞膜,分解后排出体外,从而达到解毒的目的.着重介绍了近年来谷胱甘肽硫转移酶结构与功能研究的进展,详细描述并比较了多种同工酶的三级结构、生化功能、催化机制以及底物特异性,同时对GSTs种类之间结构与功能的进化做了较深入探讨.     

关于隐形眼镜对眼睛伤害的研究

为研究隐形眼镜对人眼的伤害,笔者对隐形眼镜中的病菌进行了培养、形态学观察、革兰氏染色、生理生化试验等研究,证明隐形眼镜中含有金黄色葡萄球菌、曲霉菌等病菌,上述病菌在人的眼睛里会引起角膜炎、结膜炎等眼疾,对眼睛造成伤害。因此,笔者呼吁人们尽量少戴隐形眼镜,保护自己心灵的窗户。     

Calcitic microlenses as part of the photoreceptor system in brittlestars

Photosensitivity in most echinoderms has been attributed to 'diffuse' dermal receptors. Here we report that certain single calcite crystals used by brittlestars for skeletal construction are also a component of specialized photosensory organs, conceivably with the function of a compound eye. The analysis of arm ossicles in Ophiocoma showed that in light-sensitive species, the periphery of the labyrinthic calcitic skeleton extends into a regular array of spherical microstructures that have a characteristic double-lens design. These structures are absent in light-indifferent species. Photolithographic experiments in which a photoresist film was illuminated through the lens array showed selective exposure of the photoresist under the lens centres. These results provide experimental evidence that the microlenses are optical elements that guide and focus the light inside the tissue. The estimated focal distance (4-7 micrometer below the lenses) coincides with the location of nerve bundles-the presumed primary photoreceptors. The lens array is designed to minimize spherical aberration and birefringence and to detect light from a particular direction. The optical performance is further optimized by phototropic chromatophores that regulate the dose of illumination reaching the receptors. These structures represent an example of a multifunctional biomaterial that fulfills both mechanical and optical functions.     

间隙连接在鸡胚水晶体发育中的作用

初步报道已揭示，单克隆抗体ＮＤ６可影响鸡胚水晶体的发育，使之明显增大．ＮＤ６是一种被认为专一于膜蛋白ＭＰ２６细胞外侧段的单抗．ＭＰ２６已被认为是水晶体纤维细胞间隙连接的成份．因此，ＮＤ６很可能可阻断水晶体纤维细胞间隙连接的形成．本人试图用定量显微镜技术证实ＮＤ６对水晶体发育的影响与间隙连接数量减少之间的相关性。注射ＮＤ６于２０期鸡胚的右眼；同胚未注射的左眼作为对照．培养２４ｈ后周定，然后作下列三种处理：（１）测量整体水晶体的大小；（２）制备水晶体超薄切片，并统计间隙连接数量；（３）制备鸡胚头部的连续石蜡切片，并统计水晶体纤维细胞的数量．实验结果指出，在ＮＤ６处理２４ｈ，水晶体大小及纤维细胞数量均比对照组明显增大（表１，Ｐ＜０．０１或０．００１）；而间隙连接数量则比对照组明显减少（表２），Ｐ＜０．００１）．这表明，水晶体的增大及纤维细胞的增多是由间隙连接的减少所引起．这些结果证实，间隙连接在鸡胚水晶体的发育中起重要作用．     

Modest stabilization by most hydrogen-bonded side-chain interactions in membrane proteins

Understanding the energetics of molecular interactions is fundamental to all of the central quests of structural biology including structure prediction and design, mapping evolutionary pathways, learning how mutations cause disease, drug design, and relating structure to function. Hydrogen-bonding is widely regarded as an important force in a membrane environment because of the low dielectric constant of membranes and a lack of competition from water. Indeed, polar residue substitutions are the most common disease-causing mutations in membrane proteins. Because of limited structural information and technical challenges, however, there have been few quantitative tests of hydrogen-bond strength in the context of large membrane proteins. Here we show, by using a double-mutant cycle analysis, that the average contribution of eight interhelical side-chain hydrogen-bonding interactions throughout bacteriorhodopsin is only 0.6 kcal mol(-1). In agreement with these experiments, we find that 4% of polar atoms in the non-polar core regions of membrane proteins have no hydrogen-bond partner and the lengths of buried hydrogen bonds in soluble proteins and membrane protein transmembrane regions are statistically identical. Our results indicate that most hydrogen-bond interactions in membrane proteins are only modestly stabilizing. Weak hydrogen-bonding should be reflected in considerations of membrane protein folding, dynamics, design, evolution and function.     

鱼眼镜头光学设计

鱼眼镜头的视场高达180°或超过180°,可以作为全景电影的摄影和放映镜头,本文介绍了它的结构型式的发展,讨论了它的成象理论,给出了适合它的理想象高公式和光栏球差公式,最后提供了一个实例。     

Homologies between gap junction proteins in lens, heart and liver

The cells in the mammalian lens are electrically and metabolically coupled with each other by a network of gap junctions. These are clusters of transmembrane channels by which the fibre cells situated deeper in the lens communicate through the epithelium with the aqueous humour, the source of nutrients for the lens. Hence gap junctions are important for lens transparency. The gap junction proteins in the mammalian lens have not yet been identified with certainty. A putative fibre gap junction protein of relative molecular mass 26,000 (26K) is not related to those from other tissues, such as the liver 28K junction component. Another lens membrane protein with Mr 70K (MP70) has also been localized in the lens fibre gap junctions. Here we demonstrate by amino-terminal sequence analysis that MP70 and its in vivo-processed form, MP38 (ref. 8), belong to a wider family of gap junction proteins. With this new data on the lens, homologies between gap junction proteins now extend to organs derived from all three embryonal layers, endoderm (liver), mesoderm (heart) and ectoderm (lens).     

Thiamine derivatives bind messenger RNAs directly to regulate bacterial gene expression

Although proteins fulfil most of the requirements that biology has for structural and functional components such as enzymes and receptors, RNA can also serve in these capacities. For example, RNA has sufficient structural plasticity to form ribozyme and receptor elements that exhibit considerable enzymatic power and binding specificity. Moreover, these activities can be combined to create allosteric ribozymes that are modulated by effector molecules. It has also been proposed that certain messenger RNAs might use allosteric mechanisms to mediate regulatory responses depending on specific metabolites. We report here that mRNAs encoding enzymes involved in thiamine (vitamin B(1)) biosynthesis in Escherichia coli can bind thiamine or its pyrophosphate derivative without the need for protein cofactors. The mRNA-effector complex adopts a distinct structure that sequesters the ribosome-binding site and leads to a reduction in gene expression. This metabolite-sensing regulatory system provides an example of a 'riboswitch' whose evolutionary origin might pre-date the emergence of proteins.     

Assembly reflects evolution of protein complexes

A homomer is formed by self-interacting copies of a protein unit. This is functionally important, as in allostery, and structurally crucial because mis-assembly of homomers is implicated in disease. Homomers are widespread, with 50-70% of proteins with a known quaternary state assembling into such structures. Despite their prevalence, their role in the evolution of cellular machinery and the potential for their use in the design of new molecular machines, little is known about the mechanisms that drive formation of homomers at the level of evolution and assembly in the cell. Here we present an analysis of over 5,000 unique atomic structures and show that the quaternary structure of homomers is conserved in over 70% of protein pairs sharing as little as 30% sequence identity. Where quaternary structure is not conserved among the members of a protein family, a detailed investigation revealed well-defined evolutionary pathways by which proteins transit between different quaternary structure types. Furthermore, we show by perturbing subunit interfaces within complexes and by mass spectrometry analysis, that the (dis)assembly pathway mimics the evolutionary pathway. These data represent a molecular analogy to Haeckel's evolutionary paradigm of embryonic development, where an intermediate in the assembly of a complex represents a form that appeared in its own evolutionary history. Our model of self-assembly allows reliable prediction of evolution and assembly of a complex solely from its crystal structure.