Amyloid beta protein precursor is possibly a heparan sulfate proteoglycan core protein

The amyloid beta protein peptide is a major constituent of amyloid plaque cores in Alzheimer's disease and is apparently derived from a higher molecular weight precursor. It is now shown that the core protein of a heparan sulfate proteoglycan secreted from a nerve cell line (PC12) has an amino acid sequence and a size very similar to those of the amyloid beta protein precursor and that these molecules are antigenically related. This amyloid beta protein precursor-related protein is not found in the conditioned medium of a variant cell line (F3 PC12) that does not secrete heparan sulfate proteoglycan. The synaptic localization and metabolism of this class of proteoglycans are consistent with its potential involvement in central nervous system dysfunction.     

P1 of strawberry vein banding virus,a multilocalized protein,functions as a movement protein and interacts with the coat protein

Although the complete nucleotide sequence of strawberry vein banding virus (SVBV) has been determined and bioinformatic analysis has revealed that the SVBV genome could encode seven proteins, the precise function of each protein is unclear. This study provided evidence that the P1 protein of SVBV (SVBV-P1) possesses the following features. Bioinformatic and subcellular localization analyses showed that SVBV-P1 is localized in the cytoplasm and cell walls of epidermal cells in Nicotiana benthamiana, and it forms inclusion bodies associated with microtubules and the endoplasmic reticulum. Dilution experiments demonstrated that SVBV-P1 could move from the original agro-infiltrated cells to adjacent cells in N. benthamiana leaves. Further trans-complementation experiments demonstrated that SVBV-P1 could facilitate the intercellular movement of a movement-deficient potato virus X mutant in N. benthamiana leaves. Finally, yeast two-hybrid and bimolecular fluorescence complementation assays revealed that SVBV-P1 could interact with the SVBV coat protein, which is a major component of Caulimovirus virions. Results of the electrophoretic mobility shift assay indicated that SVBV-P1 lacks DNA-binding capability. In summary, the results suggest that SVBV-P1 is probably a movement protein of SVBV, providing new insights into the function of movement proteins of the Caulimovirus genus.     

Aldolase activity of a Plasmodium falciparum protein with protective properties

Immunization with a 41-kilodalton blood stage antigen (p41) of Plasmodium falciparum induces immunity to malaria in monkeys. However, antigenic polymorphism and repetitive amino acids commonly found in protective antigens complicate vaccine development. The gene encoding p41 has now been cloned and analyzed. Sequencing and hybridization studies revealed that the gene structure is highly conserved in 14 parasite isolates from three continents. This finding and the lack of repetitive amino acids in the translated DNA sequence may indicate that p41 has an essential function. In this study the protein was found to be 60 percent homologous to the key glycolytic enzyme aldolase from vertebrates, and the affinity-purified p41 protein from parasites showed aldolase activity.     

Tat protein from human immunodeficiency virus forms a metal-linked dimer

Tat, the transactivating protein from HIV, forms a metal-linked dimer with metal ions bridging cysteine-rich regions from each monomer. This novel arrangement is distinct from the "zinc finger" domain observed in other eukaryotic regulatory proteins. Ultraviolet absorption spectra show that Tat binds two Zn2+ or two Cd2+ ions per monomer, and electrophoresis of the Tat-metal complexes demonstrates that the protein forms metal-linked dimers. Partial proteolysis and circular dichroism spectra suggest that metal binding has its primary effects in the cysteine-rich region and relatively little effect on the folding of other regions. These results suggest new directions for biological studies and new approaches to drug design.     

Chemostimulatory protein: a new type of taste stimulus

Three taste-active proteins have recently been discovered. It is proposed that two of these (monellin and thaumatin) should be classified as chemostimulatory proteins because of their sensory effect; these two proteins taste intensely sweet. The third protein (miraculin), a taste-modifier protein, changes the normal sour taste of acids to sweet. The taste-modifier protein, miraculin, occurs in the fruit of the tropical plant Synsepalum dulcificum. Though itself not sweet, it is able to change the taste of acids from sour to sweet after the tongue has been treated with the protein. Miraculin is a basic glycoprotein with a molecular weight of 44,000. Monellin, a chemostimulatory protein, is found in the fruit of a different tropical plant, Dioscoreophyllum cumminsii. It has been characterized as a basic protein with a molecular weight of 10,700 that contains no carbohydrate. Thaumatin, another chemostimulatory protein, occurs in the fruit of a third tropical plant, Thaumatococcus daniellii. Like monellin, it is a basic protein that contains no carbohydrate. Its molecular weight is around 21,000. Certain gross similarities among the three proteins have been noted. Their basic ionic character and some features of the amino acid compositions are similar. Little is known of the structural features of the chemostimulatory proteins that are required for eliciting their intense sweetness; they are of the order of 10(5) times more effective than sucrose. The precise role of the tertiary structure in their biological activity is not known but appears to be an important area for further study. The relatively large size (11,000 to 21,000 molecular weight) of the chemostimulatory proteins provides indirect evidence that the initial interaction of these stimuli with taste receptor cells occurs at the plasma membrane.     

Molecular architecture and evolution of a modular spider silk protein gene

Spider flagelliform silk is one of the most elastic natural materials known. Extensive sequencing of spider silk genes has shown that the exons and introns of the flagelliform gene underwent intragenic concerted evolution. The intron sequences are more homogenized within a species than are the exons. This pattern can be explained by extreme mutation and recombination pressures on the internally repetitive exons. The iterated sequences within exons encode protein structures that are critical to the function of silks. Therefore, attributes that make silks exceptional biomaterials may also hinder the fixation of optimally adapted protein sequences.     

Inheritance of a serum protein in swine

A study of polymorphism of the starch-gel electrophoretic pattern for one of the blood serum proteins in swine (tentatively designated protein B) reveils that it is controlled by a single palir of alleles exhibiting partial dominance. BB genotypes appear to have twice the amount of protein B that the Bb genotype has, while bb genotypes show no evidence of the protein. Present indications are that the Yorkshire and Landrace breeds differ in the frequiency of these genes.     

Homology of beta-lactoglobulin, serum retinol-binding protein, and protein HC

The milk protein beta-lactoglobulin has been extensively studied but its function has not been identified. A clue regarding the function of a protein can be obtained by discovering a genetic relationship with a protein of known function through comparisons of amino acid sequence. Such comparisons revealed that beta-lactoglobulin is similar to human serum retinol-binding protein and to another human protein of unknown function known as complex-forming glycoprotein heterogeneous in charge (protein HC). beta-Lactoglobulins from several species have been found to bind retinol, while the absorption and fluorescence properties reported for the unidentified heterogeneous prosthetic group of protein HC are retinoid-like. The role of serum retinol-binding protein in vitamin A transport in the circulation suggests that the other two homologous proteins may function in the binding and transport of retinoids; beta-lactoglobulin may facilitate the absorption of vitamin A from milk and protein HC may mediate the excretion of retinol-derived metabolites.     

Submaxillary gland of mouse: properties of a purified protein affecting muscle tissue in vitro

A protein from the salivary gland of mice has been highly purified. It affects embryonic muscle tissue in vitro and has both esterase and peptidase activities. Addition of the pure protein to tissue culture in synthetic medium causes dissociation of muscle fibers in individual myoblasts with loss of myosin. This biological activity, as well as the esterase activity, is inhibited by low concentrations of phenylmethanesulfonyl fluoride; this suggests that the effect on the tissue is a consequence of the protein's enzymatic activities.     

Cloning and expression of a Xenopus embryonic gap junction protein

Gap junctions in the early amphibian embryo may play a fundamental role in the regulation of differentiation by mediating the cell-to-cell transfer of chemical signals. A complementary DNA encoding a gap junction present in Xenopus oocytes and early embryos has now been cloned and sequenced. This protein sequence is homologous to the well-characterized gap junction structural proteins rat connexin32 and connexin43. RNA blot analysis of total Xenopus oocyte RNA showed hybridization to a single 1.6-kilobase band. This messenger RNA is abundant in oocytes, decreases to levels below the sensitivity of our assay by stage 15 (18 hours), and is not detectable in RNA from a number of adult organs. To confirm that the oocyte cDNA encodes a gap junction channel, the protein was over expressed in Xenopus oocytes by injection of RNA synthesized in vitro. Pairs of RNA-injected oocytes formed many more time- and voltage-sensitive cell-cell channels than water-injected pairs.     

A calcium-dependent protein kinase with a regulatory domain similar to calmodulin.

Calcium can function as a second messenger through stimulation of calcium-dependent protein kinases. A protein kinase that requires calcium but not calmodulin or phospholipids for activity has been purified from soybean. The kinase itself binds calcium with high affinity. A complementary DNA clone for this kinase has been identified; it encodes a protein with a predicted molecular mass of 57,175 daltons. This protein contains a catalytic domain similar to that of calmodulin-dependent kinases and a calmodulin-like region with four calcium binding domains (EF hands). The predicted structure of this kinase explains its direct regulation via calcium binding and establishes it as a prototype for a new family of calcium-regulated protein kinases.     

Parathyroid secretion: discovery of a major calcium-dependent protein

Bovine parathyroid tissue incubated in vitro secretes a protein that is distinct from both parathyroid hormone and proparathyroid hormone and comprises about 50 percent of the total secreted protein. This protein appears to be an aggregate consisting of two or more subunits of molecular weight 70,000 as determined by polyacrylamide gel electrophoresis in sodium dodecyl sulfate. Although the function of this protein is unknown, the secretion rates of both the protein and parathyroid hormone respond in parallel to changes in the concentration of calcium in the medium.     

Measles virus matrix protein synthesized in a subacute sclerosing panencephalitis cell line

Measles virus generally produces acute illness. Rarely, however, persistent infection of brain cells occurs, resulting in a chronic and fatal neurological disease, subacute sclerosing panencephalitis (SSPE). Evidence indicates that expression of the measles virus matrix protein is selectively restricted in this persistent infection, but the mechanism underlying this restriction has not been identified. Defective translation of matrix messenger RNA has been described in one SSPE cell line. This report presents evidence that in a different SSPE tissue culture cell line IP-3-Ca, the matrix protein is synthesized but fails to accumulate. A general scheme is proposed to reconcile the different levels at which restriction of matrix protein has been observed.     

抗冻蛋白的研究进展及其应用

抗冻蛋白自 196 4年发现以来 ,就引起了各国学者的广泛重视。它具有特殊的热滞活性 ,即使溶液的冰点低于熔点 ;它还能抑制冰晶的生长 ,特别是在较低的浓度下就有较高的重晶化抑制活性 ,保护生物免受冻害的影响。人们正试图分离和纯化高活性的抗冻蛋白 ,以应用于组织、细胞、胚胎和器官的低温保存 ;同时还进行抗冻蛋白基因工程的研究 ,期望能够改良某些生物的抗冻性能。介绍了已知抗冻蛋白的结构、抗冻机制和抗冻作用 ,并讨论了抗冻蛋白基因工程的研究进展及抗冻蛋白的应用前景     

Selective inhibition of a regulatory subunit of protein phosphatase 1 restores proteostasis

Many biological processes are regulated through the selective dephosphorylation of proteins. Protein serine-threonine phosphatases are assembled from catalytic subunits bound to diverse regulatory subunits that provide substrate specificity and subcellular localization. We describe a small molecule, guanabenz, that bound to a regulatory subunit of protein phosphatase 1, PPP1R15A/GADD34, selectively disrupting the stress-induced dephosphorylation of the α subunit of translation initiation factor 2 (eIF2α). Without affecting the related PPP1R15B-phosphatase complex and constitutive protein synthesis, guanabenz prolonged eIF2α phosphorylation in human stressed cells, adjusting the protein production rates to levels manageable by available chaperones. This favored protein folding and thereby rescued cells from protein misfolding stress. Thus, regulatory subunits of phosphatases are drug targets, a property used here to restore proteostasis in stressed cells.     

Microtubule-associated protein MAP2 shares a microtubule binding motif with tau protein

The microtubule-associated protein MAP2 is a prominent large-sized component of purified brain microtubules that, like the 36- to 38-kilodalton tau proteins, bears antigenic determinants found in association with the neurofibrillary tangles of Alzheimer's disease. The complete sequence of mouse brain MAP2 was determined from a series of overlapping cloned complementary DNAs. The sequence of the carboxyl-terminal 185 amino acids is very similar (67 percent) to a corresponding region of tau protein, and includes a series of three imperfect repeats, each 18 amino acids long and separated by 13 or 14 amino acids. A subcloned fragment spanning the first two of the 18-amino acid repeats was expressed as a polypeptide by translation in vitro. This polypeptide copurified with microtubules through two successive cycles of polymerization and depolymerization, whereas a control polypeptide derived from the amino-terminal region of MAP2 completely failed to copurify. These data imply that the carboxyl-terminal domain containing the 18-amino acid repeats constitutes the microtubule binding site in MAP2. The occurrence of these repeats in tau protein suggests that these may be a general feature of microtubule binding proteins.     

蛋白质交联与食品蛋白配料功能性的研究进展

为研究蛋白质交联技术对蛋白质的功能性作用,对国内外食品体系中蛋白质交联的研究状况进行了系统回顾。首先,围绕食品蛋白配料最重要的两种功能性(乳化性、凝胶性),分析了"蛋白-蛋白"与"蛋白-糖"两种交联类型各自对乳化功能及凝胶功能的促进作用;其次,进一步探讨了不同蛋白质交联对热稳定性、亲水性、成膜性、发泡性和其他食品配料的常见加工性能的影响。研究表明:多数与蛋白质有关的交联对食品蛋白配料功能均产生积极作用,且针对"蛋白-蛋白"交联的研究多于"蛋白-糖"交联;国内外有关蛋白质交联的研究涵盖了动物、植物、乳和蛋等主要食品蛋白源,且多数集中在对单一蛋白源的研究,而对混合蛋白源的研究相对匮乏;今后应针对蛋白质交联机理,特别是加工过程中蛋白自然氧化进行深入探索,将蛋白源应用范围进一步扩展到低值蛋白、加工副产物及混合蛋白源。     

A cytoplasmic protein inhibits the GTPase activity of H-Ras in a phospholipid-dependent manner

A cytoplasmic protein has been identified that inhibits the guanosine triphosphatase (GTPase) activity of bacterially synthesized, cellular H-Ras protein. This GTPase inhibiting protein is able to counteract the activity of GTPase activating protein (GAP), which has been postulated to function as a negative regulator of Ras activity. The potential biological importance of the GTPase inhibiting protein is further supported by its interaction with lipids. Phospholipids produced in cells as a consequence of mitogenic stimulation increase the activity of the GTPase inhibiting protein, as well as inhibit the activity of GAP. The interaction of such lipids with each of these two regulatory proteins would, therefore, tend to increase the biological activity of Ras and stimulate cell proliferation.     

NMR structure of Mistic, a membrane-integrating protein for membrane protein expression

Although structure determination of soluble proteins has become routine, our understanding of membrane proteins has been limited by experimental bottlenecks in obtaining both sufficient yields of protein and ordered crystals. Mistic is an unusual Bacillus subtilis integral membrane protein that folds autonomously into the membrane, bypassing the cellular translocon machinery. Using paramagnetic probes, we determined by nuclear magnetic resonance (NMR) spectroscopy that the protein forms a helical bundle with a surprisingly polar lipid-facing surface. Additional experiments suggest that Mistic can be used for high-level production of other membrane proteins in their native conformations, including many eukaryotic proteins that have previously been intractable to bacterial expression.     

Long-range interactions within a nonnative protein

Protein folding and unfolding are coupled to a range of biological phenomena, from the regulation of cellular activity to the onset of neurodegenerative diseases. Defining the nature of the conformations sampled in nonnative proteins is crucial for understanding the origins of such phenomena. We have used a combination of nuclear magnetic resonance (NMR) spectroscopy and site-directed mutagenesis to study unfolded states of the protein lysozyme. Extensive clusters of hydrophobic structure exist within the wild-type protein even under strongly denaturing conditions. These clusters involve distinct regions of the sequence but are all disrupted by a single point mutation that replaced residue Trp62 with Gly located at the interface of the two major structural domains in the native state. Thus, nativelike structure in the denatured protein is stabilized by the involvement of Trp62 in nonnative and long-range interactions.