Immunochemical studies on barley seed storage proteins

The antigenic relationships between the prolamins of barley, rye and wheat have been studied by examining the specificity of an antibody to C hordein in a quantitative study using a laser nephelometer. The antibody reacts weakly with B hordein and strongly with 75-kdalton and 40-kdalton -secalins from rye and ₃ some -gliadins from wheat. Absorption experiments and immunodiffusion tests indicate that there are shared antigenic determinants for most of the prolamins. All the species with reacting prolamins belong to the sub-family Festucoideae of the Gramineae. The prolamins of maize, pearl millet and sorghum, species of the sub-family Panicoideae, do not react. The results confirm the known lack of homology between the prolamins of the two sub-families and also indicate the presence of relationships not yet established between C hordein, the 75-kdalton and 40-kdalton -secalins and also ₃ gliadin.Abbreviations HMW high molecular weight - PAGE polyacnylamide-gel electrophoresis - PBS phosphate-buffered saline - RLS relative light scattering - SDS sodlum dodecyl sulphate     

Immunochemical identification and partial characterization of female-specific serum proteins in white-edged rockfish,Sebastes taczanowskii

Synopsis Female-specific serum proteins (FSSPs) in white-edged rockfish,Sebastes taczanowskii, were identified and partially characterized by immunochemical procedures. Two FSSPs, which clearly reacted with antiserum against egg proteins, were confirmed in the serum of mature females, and estrogen treatment induced similar FSSPs in the serum of mature males. Hence, the FSSPs were considered to be vitellogenin. The vitellogenin concentration in female fish was high during the vitellogenic period and low during gestation, parturition and the recovery period, indicating that vitellogenin is used only for yolk formation in the oocytes and not as a direct nutritional source for developing embryos during gestation. On the other hand, an FSSP (FS3), which was considered not to be vitellogenin, was also identified in the sera of mature females and males after estradiol-17β administration by using an antiserum (a-FS3) that removed the components of the male serum and egg extracts from the anti-mature female serum antiserum. Moreover, immunohistochemical observation with a-FS3 illustrated that FS3 was a major constituent of the ovarian fluid but not of vitellogenic oocytes. The cross-reactivities of these FSSPs among seven viviparous rockfishes demonstrated that vitellogenin existed in the sera of all rockfishes studied belonging to the generaSebastes andSebastiscus, whereas FS3 was not present in several species ofSebastes.     

Immunochemical studies of fragments of bovine serum albumin.

Antigenic properties of 14 fragments of bovine albumin were measured using antisera to albumin and to two of its fragments. All seven fragments larger than 21,000 daltons formed immune precipitates. Although immune precipitates were not formed with smaller fragments, inhibition tests indicated the presence of antigenic sites on several of these fragments as well. The results predict the occurrence of six or more antigenic determinants and allow assignment of their positions in the parent molecule. These sites are distributed along the entire protein chain, with the sites of greatest antibody affinity situated in the COOH-terminal region. Evidence is presented that some sites are homologous, reacting with the same populations of antibodies, and that other sites are unique, binding to an exclusive population of antibodies.     

Immunochemical studies on dextran-specific and levan-specific myeloma proteins from nzb mice

Two dextran-specific (PC 3858 and PC 3936) and one levan-specific (PC 3660) NZB myeloma proteins were studied by quantitative precipitin and precipitin-inhibition assays. Both myeloma antidextrans were α-D(1→6) specific and precipitated strongly with a synthetic, linear dextran molecular weight 36,500, and with other dextrans. The two myeloma antidextrans differed with respect to their relative reactivities with dextrans containing various proportions of α-D-(1→6), α-D-(1→4)-like, and α-D-(1→3)-like linkages. In inhibition assays, the two antidextran myeloma proteins behaved differently from each other, from α-D-(1→6)-specific BALB/c myeloma antidextrans, and from the human antidextrans previously studied. Isomalto-oligosaccharides IM3, IM4, and IM5 were all equal in inhibitory power but were only about 60% as potent as IM6 and IM7, which also inhibited equally on a molar basis. Although precipitation with linear dextran suggests that both may have groove-type sites, as previously inferred for QUPC 52, the size of their combining sites is uncertain. It is not clear whether the sites are only as big as three glucose residues with the increased inhibition by six and seven glucose residues being attributable to partial bivalence and to their ability to combine in several ways along the chain, or whether the site is as big as six glucose residues with the increment in binding by the fourth and fifth glucose residues being minimal and the sixth contributing considerable additional binding-energy. The fructan-specific myeloma protein did not react with inulin, but reacted with many levans and with perennial rye-grass levan containing only β-D-(2→6) links. The levan-antilevan reaction was not inhibited by β-D-(2→1)-linked oligosaccharides. The findings suggest that PC 3660 has a specificity for (2→6)-linked chains.     

Immunochemical studies of high mobility group non-histone chromatin proteins HMG 1 and HMG 2

Sera were raised to non-histone chromatin proteins HMG 1 and HMG 2. Immunoperoxidase staining localised these proteins on chromosomes during mitosis and indicated a cell cycle-related variation in these proteins during interphase. Some species differences in HMG 1 and HMG 2 were also observed.