Immunochemical comparison of histidine-rich protein in keratohyalin granules and cornified cells.

(1) Combination of techniques for extraction and purification of histidine rich protein established by several investigators were employed for comparison of histidine-rich protein in granular cells and cornified cells of newborn rats. (2) Histidine-rich protein extracted from the same cell fraction by two different techniques either in 1 M potassium phosphate buffer (Ugel) or in 4 M urea (Dale) showed identical elution profiles on CM 52 cellulose ion exchange chromatography and the same SDS polyacrylamide gel electrophoretic patterns. (3) Histidine-rich protein from granular cells contained polypeptides of larger molecular sizes than those in histidine-rich protein from cornfield cells, although amino acid composition of the two histidine-rich protein was non-distinguishable (histidine residue was more than 7%). (4) Antibodies raised in rabbits by injection of histidine rich protein from granular cells and that from cornfield cells immunologically cross-reacted. Furthermore, the antisera were found to be reactive over both keratohyalin granules and cornified cells, but not epidermal cells of the lower strata.     

A-T rich sequences in vertebrate DNA

Partially denatured DNAs from mouse, cow, and chicken were visualized in the electron microscope by the basic protein film technique and the size and distribution of the denatured regions characterized. A-T rich sequences visualized at 15% denaturation average about 1500 bases in length for all three species and are arranged quite non-randomly in the genome. This arrangement is such that 30–50% of the entire genome contains no A-T rich DNA, and another 20% is composed about one-half of A-T rich sequences and one-half of other sequences. Comparison with DNA denaturation profiles indicates that for each organism these sequences are from 25–35% G+C and that there is very little if any DNA more A-T rich than these. Estimates from published studies of fluorescence enhancement of quinacrine bound to A-T rich DNAs suggest that the observed non-random organization of A-T rich sequences is sufficient to account for Q banding of metaphase chromosomes.     

Characterization of a camel milk protein rich in proline identifies a new beta-casein fragment

A camel milk whey protein has been isolated by reverse-phase high performance liquid chromatography. The protein is, like caseins, rich in proline (25% of the whole protein). The N-terminal amino acid sequence shows that the protein is homologous with a C-terminal region of beta-caseins analyzed from other species. The protein is concluded to be a fragment of beta-casein, derived from a non-tryptic type of cleavage of the parent molecule, and increasing the multiplicity of known casein products.     

Single cell protein production by photosynthetic bacteria grown on the clarified effluents of biogas plant

Summary Anaerobically digested cow dung was separated by centrifugation into solid residue and liquid supernatant fractions. Clarified supernatant fraction, rich in volatile fatty acids, supported the growth of photosynthetic bacteria. Single cell protein from different photosynthetic bacteria, grown on clarified supernatant, was found to be rich in essential and sulphur amino acids. Rhodopseudomonas capsulata produced the best single cell protein.     

Linkers in the structural biology of protein–protein interactions

Linkers or spacers are short amino acid sequences created in nature to separate multiple domains in a single protein. Most of them are rigid and function to prohibit unwanted interactions between the discrete domains. However, Gly‐rich linkers are flexible, connecting various domains in a single protein without interfering with the function of each domain. The advent of recombinant DNA technology made it possible to fuse two interacting partners with the introduction of artificial linkers. Often, independent proteins may not exist as stable or structured proteins until they interact with their binding partner, following which they gain stability and the essential structural elements. Gly‐rich linkers have been proven useful for these types of unstable interactions, particularly where the interaction is weak and transient, by creating a covalent link between the proteins to form a stable protein–protein complex. Gly‐rich linkers are also employed to form stable covalently linked dimers, and to connect two independent domains that create a ligand‐binding site or recognition sequence. The lengths of linkers vary from 2 to 31 amino acids, optimized for each condition so that the linker does not impose any constraints on the conformation or interactions of the linked partners. Various structures of covalently linked protein complexes have been described using X‐ray crystallography, nuclear magnetic resonance and cryo‐electron microscopy techniques. In this review, we evaluate several structural studies where linkers have been used to improve protein quality, to produce stable protein–protein complexes, and to obtain protein dimers.     

Involvement of protein disulphides and sulphydryls in chromosome banding

G-banding of chromosomes appears to be a consequence of a varying concentration of protein disulphides and sulphydryls along the chromosomes. Dark bands seem to be relatively rich in disulphides, and pale bands relatively rich in sulphydryls.     

Desmosine and isodesmosine as cross-links in the hinge-ligament protein of bivalves. 3,3'-Methylenebistyrosine as an artefact

Desmosine and isodesmosine were detected in an invertebrate molluscan species, i.e. in an insoluble protein in the hinge ligament of a bivalve species, Sakhalin surf clam (Pseudocardium sachalinensis, in family Mactridae). The protein is rich in glycine and methionine S-oxide but devoid of hydroxyproline and hydroxylysine. 3,3'-Methylenebistyrosine was also detected in the HCl hydrolysate of the hinge-ligament protein, but it was found to be an artefact produced from tyrosine and formaldehyde derived from methionine S-oxide during the HCl hydrolysis of the protein.     

Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease

Intact ribonucleic acid (RNA) has been prepared from tissues rich in ribonuclease such as the rat pancreas by efficient homogenization in a 4 M solution of the potent protein denaturant guanidinium thiocyanate plus 0.1 M 2-mercaptoethanol to break protein disulfide bonds. The RNA was isolated free of protein by ethanol precipitation or by sedimentation through cesium chloride. Rat pancreas RNA obtained by these means has been used as a source for the purification of alpha-amylase messenger ribonucleic acid.     

祁连山区几种草地蝗虫蛋白质营养评价

为评价草地蝗虫作为家禽蛋白源饲料的营养价值.本研究以几种草地蝗虫为供试对象,在分析测定其蛋白和氨基酸含量基础上,采用国际通用蛋白质营养评价标准,分析评价其蛋白质营养价值.结果表明,草地蝗虫蛋白含量高(62.4%～67.3%,DM),氨基酸组分齐全、家禽所需必需氨基酸约占蝗虫总干重的30.3%～34.2%,其氨基酸模式优...     

Molecular characterization of a surface protein antigen gene from serotype c Streptococcus mutans, implicated in dental caries

The complete nucleotide sequence of the gene for a cell-surface protein antigen (PAc) of Streptococcus mutans MT8148 (serotype c) was determined. The pac gene consisted of 4695 bp and coded for a 170773D protein. The pac gene product contained a putative 38 amino acid signal peptide, resulting in a 166817D mature protein. A potential promoter sequence and a putative Shine-Dalgarno sequence preceded the open reading frame. Two internal repeating amino acid sequences were present in the PAc. One repeating region located in the N-terminal region was rich in alanine, and the other located in the central region was rich in proline. Southern blot analysis under the less stringent condition (allowing up to 35% base mismatch) revealed that the probe covering the proline-rich region hybridized to DNA preparations from strains of Streptococcus cricetus, Streptococcus sobrinus and Streptococcus downei as well as Streptococcus mutans.     

Regulation of transcription of katE and katF in Escherichia coli

Fusion plasmids with lacZ under the control of the katE (encoding catalase or hydroperoxidase HPII) and katF (encoding a sigma factor-like protein required for katE expression) promoters were constructed. Expression from both katE and katF promoters was low in rich medium but elevated in poor medium during log-phase growth. Furthermore, the slowdown in growth as cells entered the stationary phase in rich medium, a result of carbon source depletion, was associated with an increase in katE and katF expression. A simple reduction in the carbon source level as the cells entered the stationary phase was not responsible for the increase in expression, because transferring the culture to a medium with no glucose did not induce expression from either promoter. Spent rich medium from stationary-phase cells was capable of inducing expression, as were simple aromatic acids such as benzoate, o-hydroxybenzoate, and p-aminobenzoate added to new medium. Anaerobiosis did not cause an increase in expression, nor did it significantly change the pattern of expression. Regardless of the medium, katF expression was always turned on before or coincidently with katE expression; in the presence of benzoate katF was fully induced, whereas katE was only partially induced, suggesting that a factor in addition to KatF protein was involved in katE expression. During prolonged aerobic incubation, cells lacking katF died off more rapidly than did cells lacking either katE or katG.     

Isolation of sheep anterior pituitary messenger RNA and its translation in a heterologous cell-free system

A preparation rich in the specific messenger RNA involved in the synthesis of prolactin from sheep anterior pituitary glands was obtained by employing both the immunochemical and affinity techniques. A dose-dependent and efficient stimulation of protein synthesis by the isolated total pituitary RNA as well as poly (A) rich RNA were achieved with the reticulocyte system. The synthesis of prolactin as one of the translational products of this cell-free system was established by specific immunoprecipitation followed by resolution on sodium dodecyl sulphate polyacrylamide gel electrophoresis.     

Secretion and movement of wingless protein in the epidermis of the Drosophila embryo.

The segment polarity gene wingless encodes a cysteine rich protein which is essential for pattern formation in Drosophila. Using polyclonal antibodies against the product of the wingless gene, we demonstrate that this protein is secreted in the embryo and that it is taken up by neighbouring cells. The protein can be found two or three cell diameters away from the cells in which it is synthesized. We discuss the possible mechanisms which are responsible for this spatial distribution and its regulation during embryogenesis.     

On the localization of polyribosomes in the system of chloroplast lamellae

From chloroplasts of 10-day-old pea seedlings exposed to the light for 19 h, two fractions have been isolated. One of them is rich in lamellae of the stroma, and the other is rich in thylakoids and fragments of the grana. These fractions have been obtained after centrifugation of chloroplasts disrupted by osmotic shock in a discontinuous sucrose gradient. The fraction containing thylakoids of grana differs from the fraction of lamellae of the stroma in its higher content of RNA and DNA as related to protein and in the capacity to incorporate intensively ¹⁴C amino acids into proteins. After its treatment with detergents (0.5% sodium deoxycholate and 0.4% Triton X-100) and repeated centrifugation in the discontinuous sucrose gradient it dissociates further into two fractions. During electron microscopic studies one of these fractions displays partially disrupted grana and the other exhibits extensive networks of polyribosomes incompletely liberated from proteins, including the de novo synthesized protein.The similar treatment of the fraction rich in lamellae of the stroma does not result in the liberation of polyribosomes.It is concluded that in this stage of chloroplast development, polyribosomes occurring in the lamellae system are localized in the thylakoids of grana and are not bound to lamellae of the stroma.     

The structure of the rat liver glycogen backbone protein

Rat liver glycogen was freed of non-covalently bound protein. The backbone protein was purified from the pure glycogen. This protein had a molecular weight of 60,000 daltons and amino acid analysis showed it to be rich in glutamate, serine and the hydrophobic amino acids. In both size and amino acid content it differed from the corresponding rabbit muscle protein. An O-type glycoprotein linkage is suggested.     

番茄花粉特异表达的高赖氨酸蛋白基因(tsb)的克隆与特性分析

赖氨酸是人和单胃动物必需的八种氨基酸之一,食物蛋白中的必需氨基酸种类不全或数量不足,都会影响其它氨基酸的利用.玉米等主要禾谷类作物由于缺少赖氨酸,种子蛋白利用率还不到50%.在长期以玉米为口粮的地区,人们往往患有营养不足或糙皮病.     

Deglycosylation by gaseous hydrogen fluoride of mucus glycoproteins immobilized on nylon membranes and in microtiter wells

Strongly reacting antibodies specific for defined mucin gene products are often directed against the mucin protein backbone of the heavily glycosylated serine/threonine rich regions. A prerequisite for the use of such antibodies is often the complete removal of the oligosaccharides from the protein. This paper describes an efficient one-step deglycosylation method using gaseous hydrogen fluoride on nylon blotting membranes and microtiter wells.     

Metabolic engineering of Escherichia coli for the production of medium-chain-length polyhydroxyalkanoates rich in specific monomers

The Escherichia coli fabG(Ec) gene and the Pseudomonas aeruginosa rhlG(Pa) gene, which encode 3-ketoacyl-acyl carrier protein reductase, were expressed in E. coli W3110 and its fadA mutant strain WA101 to examine their roles in medium-chain-length (MCL) polyhydroxyalkanoate (PHA) biosynthesis from fatty acids. When one of these 3-ketoacyl-acyl carrier protein reductase genes was co-expressed with the Pseudomonas sp. 61-3 PHA synthase gene (phaC2(Ps)) in E. coli W3110, MCL-PHA composed mainly of 3-hydroxyoctanoate and 3-hydroxydecanoate was synthesized from sodium decanoate. When the fabG(Ec) gene and the phaC2(Ps) gene were co-expressed in the fadA mutant E. coli strain WA101, MCL-PHA rich in 3-hydroxydecanoate monomer up to 93 mol% was accumulated from sodium decanoate. This was possible by efficiently redirecting 3-ketoacyl-coenzymes A from the beta-oxidation pathway to the PHA biosynthesis pathway without losing two carbon units, the strategy of which can be extended for the production of MCL-PHAs rich in other specific monomers.