Protein and free amino acids in wheat grain as affected by soil types and salinity levels in irrigation water

Summary The effects of four lysimeter soil series under three salinity levels were evaluated for grain yield, wt/1000 seeds, protein, and amino acids in Mexican dwarf wheat (Triticum aestivum L. var. Cajeme 71). The soil series consisted of: Holtville clay loam, Greenfield sandy loam, San Emigdio sandy loam, and Altamont clay loam. The irrigation water salinity levels were designated: low –2.2 mmho, medium –4.2 mmho, and high –7.1 mmho.No significant differences were found in the amount of grain harvested or wt/1000 seeds in the 1976 crop produced on the differential soil series. The yield of the 1977 crop was significantly affected by the soil types.Effects of soil type on the protein amino acids in the grain in both years were similar. Significantly higher protein amino acid levels of histidine, arginine, aspartic acid, threonine, serine, glutamic acid, glycine, alanine, cystine, valine, methionine, isoleucine, leucine, tyrosine, and phenylalanine were found in the grain grown on Altamont clay loam soil than the other types.The free amino acids in grain from the 1976 and 1977 crops were similarly affected by the soil types, except that the quantitative values of the free amino acids were substantially lower in 1977 than in 1976. The free amino acids significantly influenced by soil types were tryptophane, lysine, arginine, aspartic acid, threonine, serine, glycine, alanine, valine, isoleucine, tyrosine, and phenylalanine. In both years' crops, the sum of the free amino acid fractions was significantly higher in the grain produced on the Altamont soil than on the other soils.Salinity level in the irrigation water did not affect the 1976 crop yield or wt/1000 seeds. Although yields of the 1977 crop were significantly reduced by salinity, the wt/1000 seeds was not. The sum of protein amino acids was significantly higher in the 1976 and 1977 grain crops irrigated with high salinity water than in low salinity irrigated crops.An increased salinity irrigation water significantly reduced the sum of free amino acid fractions in the 1976 grain crop. Since some of the free amino acids in the 1977 grain crop increased while the others decreased due to the salinity level in the irrigation water, the sum of the free amino acid fractions was not significantly influenced.Significant interactions were found between soil types and salinity levels on free arginine, threonine, serine, glutamic acid, and alanine, and also on the sum of the free amino acids in the 1976 wheat grain. In the 1977 wheat grain, there were significant interactions between soil types and salinity levels on the free glutamic acid, valine, leucine, tyrosine, and phenylalanine, and on protein serine, glutamic acid, glycine, alanine, and the sum of the protein amino acids.The amounts of essential amino acids expressed as mg of amino acid/g of protein were not affected by the soil types or salinity levels. With the exception of lysine, and possibly threonine and methionine plus cystine, the essential amino acids were present in the grain at concentrations equal to or greater than recommended by WHO and FAO.     

Inactivation and reactivation of proteins (enzymes)

The molecular mechanisms of protein inactivation, i.e. aggregation, thiol-disulphide exchange, alteration of the primary structure, dissociation of cofactor molecules from the active centre, dissociation of the oligomeric proteins into subunits and conformational changes have been analysed. All these mechanisms are closely interrelated during inactivation of proteins. However, in many cases, the conformational changes accompany and trigger other inactivation processes. Reactivation of irreversibly inactivated proteins is·discussed. Reactivation can be successful when inactivation has been caused by aggregation, modification of SH-groups (or S-S bonds) or as a consequence of irreversible conformational changes.     

Nature of amino acid substitutions in homologous proteins during evolution

Replacement substitutions of mitochondrial cytochrome $\text{c}$ and α- and β-chains of haemoglobin have been studied by considering the structural similarity among amino acid residues at the secondary and tertiary structural levels. Secondary structural similarity explains ～70% while tertiary structural similarity explains ～50% of observed replacements for most of the cases. These structural similarities could not account for all the replacement substitutions. The study was extended to consider the composition of codons, and the chemical nature and polarity of the replacing and replaced residues. These also could not individually account for all the affected replacements. In general, no property of amino acid residues is conserved for substitutions occurring at any single position during evolution of proteins.     

A structural model for the chromophore-binding domain of ovine rhodopsin

A putative model for the structure of the relatively independent carboxyl-terminal domain of (rhod)opsin has been developed by use of a combination of several secondary structure prediction methods. The validity of this approach was confirmed by comparing the secondary structure for bacteriorhodopsin as predicted by these methods with its known low resolution structure. The resulting predicted structure agreed well with the experimental data. The model obtained for opsin incorporates two transmembrane α-helical rods linked by an intradiscal loop. Each of the helical sections is interrupted by a short irregular region. One of these includes the lysyl residue to which the chromophore 11-cis retinal is attached. The second non-regular segment, almost opposite the first, contains a cysteinyl and a tryptophanyl residue which may be involved in protein—chromophore interaction. The proposed structure of this whole domain could prove instructive in the elucidation of the primary events of visual transduction.     

Preservation of proteins in mummified tissues

Protein material was extracted from the dessicated tissues of several Egyptian mummies and a frozen Eskimo. The distribution and degree of preservation of high molecular weight protein was analyzed by gel filtration, protein assays, amino acid analysis, and polyacrylamide gel electrophoresis. The protein has undergone considerable degradation although some high molecular weight protein (C. 130,000 daltons) remains intact. Amino acid analysis of the extracted protein indicates the basic amino acids have undergone a chemical modification and may represent a point of preferential breakdown in the polypeptide chain. Atomic absorption spectrophotometry of tissue cations suggests a correlation between degree of preservation of mummified tissue and levels of sodium salts (natron) in the tissue.     

Cortactin Controls Surface Expression of the Voltage-gated Potassium Channel KV10.1

K_V10.1 is a voltage-gated potassium channel aberrantly expressed in many cases of cancer, and participates in cancer initiation and tumor progression. Its action as an oncoprotein can be inhibited by a functional monoclonal antibody, indicating a role for channels located at the plasma membrane, accessible to the antibody. Cortactin is an actin-interacting protein implicated in cytoskeletal architecture and often amplified in several types of cancer. In this study, we describe a physical and functional interaction between cortactin and K_V10.1. Binding of these two proteins occurs between the C terminus of K_V10.1 and the proline-rich domain of cortactin, regions targeted by many post-translational modifications. This interaction is specific for K_V10.1 and does not occur with K_V10.2. Cortactin controls the abundance of K_V10.1 at the plasma membrane and is required for functional expression of K_V10.1 channels.     

Chemical composition and stability of Nb1-Particles fromNitrobacter agilis

Nb₁-particles fromNitrobacter agilis were found to be highly stable and could only be disrupted by chemicals or prolonged sonication.Spectra of the Nb₁-particles indicated that protein is their major component. They contain no lipid.Highly purified Nb₁-particles that were electronmicroscopically free from contaminating membranes, contained 7 different proteins, as shown by sodium dodecyl sulfate-polyacrylamide gelelectrophoresis.Abbreviations SDS sodium dodecyl sulfate - PAGE polyacrylamide-gelelectrophoresis - M. W. molecular weight - O.D. opitical density - HAA hepatitis associated antigen     

Analysis of the developmental expression of small VCP-interacting protein and its interaction with steroidogenic acute regulatory protein in Leydig cells

Small VCP-interacting protein (SVIP) is a 9-kDa protein that is composed of 76 amino acids, and it plays a role in the endoplasmic reticulum-associated protein degradation (ERAD) pathway. Recent studies have shown that SVIP is an androgen-responsive protein and its expression is regulated by androgens. Because no data are available regarding the cellular localization and expression of SVIP in the mouse testis, where androgens are highly expressed, immunohistochemistry and western blotting were performed. In the fetal testis, we found that moderate but consistent staining of SVIP is present in the cytoplasm of Leydig cells. In prepubertal and adult life, SVIP remains present in Leydig cells as well as in the cytoplasm of some peritubular and Sertoli cells. From postnatal day 15 onward, SVIP is strongly expressed in the cytoplasm of Leydig cells.Furthermore, TM3, MA-10 Leydig and Sertoli cell lines were also used to evaluate the expression of SVIP. To identify the interacting partners, such as steroidogenic acute regulatory (STAR) protein, colocalization studies were performed by fluorescence microscopy, showing that STAR colocalized with SVIP in the adult mouse testis. The expression changes of STAR were studied by using SVIP siRNAs in Leydig cell line cultures. Depletion of SVIP resulted in decreased expression of STAR. Additionally, the number and size of lipid droplets were significantly increased in SVIP-depleted Leydig cells. Taken together, our data identify SVIP as a marker of Leydig cell lineage and as a regulator of STAR protein expression and lipid droplet status in Leydig cells.     

Covalent Binding of BMP-2 on Surfaces Using a Self-assembled Monolayer Approach

Bone morphogenetic protein 2 (BMP-2) is a growth factor embedded in the extracellular matrix of bone tissue. BMP-2 acts as trigger of mesenchymal cell differentiation into osteoblasts, thus stimulating healing and de novo bone formation. The clinical use of recombinant human BMP-2 (rhBMP-2) in conjunction with scaffolds has raised recent controversies, based on the mode of presentation and the amount to be delivered. The protocol presented here provides a simple and efficient way to deliver BMP-2 for in vitro studies on cells. We describe how to form a self-assembled monolayer consisting of a heterobifunctional linker, and show the subsequent binding step to obtain covalent immobilization of rhBMP-2. With this approach it is possible to achieve a sustained presentation of BMP-2 while maintaining the biological activity of the protein. In fact, the surface immobilization of BMP-2 allows targeted investigations by preventing unspecific adsorption, while reducing the amount of growth factor and, most notably, hindering uncontrolled release from the surface. Both short- and long-term signaling events triggered by BMP-2 are taking place when cells are exposed to surfaces presenting covalently immobilized rhBMP-2, making this approach suitable for in vitro studies on cell responses to BMP-2 stimulation.