Acid Polyacrylamide Gel Electrophoresis Research Articles

Analysis of the Native Quaternary Structure of Vanilloid Receptor 1

Vanilloid receptor subtype 1 (VR1) is a ligand-gated channel that can be activated by capsaicin and other vanilloids as well as by protons and heat. In the present study, we have analyzed the oligomeric state of VR1. Co-immunoprecipitation of differently tagged VR1 molecules indicated that VR1 can form oligomers. Using two different heterologous VR1 expression systems as well as endogenous VR1 expressed in dorsal root ganglion cells, we analyzed oligomer formation using perfluoro-octanoic acid polyacrylamide gel electrophoresis. Results were confirmed both with chemical cross-linking agents as well as through endogenous cross-linking mediated by transglutaminase. Our results clearly show that VR1 forms multimers in each of the expression systems with a homotetramer as a predominant form. The oligomeric structure of VR1 may contribute to the complexity of VR1 pharmacology. Finally, differences in glycosylation between the systems were observed, indicating the need for caution in the use of the heterologous expression systems for analysis of VR1 properties.

Dopamine β-Monooxygenase Signal/Anchor Sequence Alters Trafficking of Peptidylglycine α-Hydroxylating Monooxygenase

Dopamine beta-monooxygenase (DBM) and peptidylglycine alpha-hydroxylating monooxygenase (PHM) are essential for the biosynthesis of catecholamines and amidated peptides, respectively. The enzymes share a conserved catalytic core. We studied the role of the DBM signal sequence by appending it to soluble PHM (PHMs) and expressing the DBMsignal/PHMs chimera in AtT-20 and Chinese hamster ovary cells. PHMs produced as part of DBMsignal/PHMs was active. In vitro translated and cellular DBMsignal/PHMs had similar masses, indicating that the DBM signal was not removed. DBMsignal/PHMs was membrane-associated and had the properties of an intrinsic membrane protein. After in vitro translation in the presence of microsomal membranes, trypsin treatment removed 2 kDa from DBMsignal/PHMs while PHMs was entirely protected. In addition, a Cys residue in DBMsignal/PHMs was accessible to Cys-directed biotinylation. Thus the chimera adopts the topology of a type II membrane protein. Pulse-chase experiments indicate that DBMsignal/PHMs turns over rapidly after exiting the trans-Golgi network. Although PHMs is efficiently localized to secretory granules, DBMsignal/PHMs is largely localized to the endoplasmic reticulum in AtT-20 cells. On the basis of stimulated secretion, the small amount of PHMs generated is stored in secretory granules. In contrast, the expression of DBMsignal/PHMs in PC12 cells yields protein that is localized to secretory granules.

Connexin45 Interacts with Zonula Occludens-1 and Connexin43 in Osteoblastic Cells

The relative expression of connexin43 and connexin45 modulates gap junctional communication and production of bone matrix proteins in osteoblastic cells. It is likely that changes in gap junction permeability are determined by the interaction between these two proteins. Cx43 interacts with ZO-1, which may be involved in trafficking of Cx43 or facilitating interactions between Cx43 and other proteins. In this study we sought to identify proteins that associate with Cx45 by coprecipitation in non-denaturing conditions. Cx45 was isolated with a 220-kDa protein that we identified as ZO-1. Under the same conditions, Cx43 also was isolated with anti-Cx45 antiserum from Cx45-transfected ROS cells (ROS/Cx45 cells). Cx43 antiserum could also coprecipitate ZO-1 in the transfected and untransfected ROS cells. Double label immunofluorescence studies showed that ZO-1, Cx43, and Cx45 colocalized at appositional membranes in ROS/Cx45 cells suggesting that all three proteins are normally associated in the cells. Additionally, we found that in vitro translated ZO-1 binds to the carboxyl-terminal of Cx45 indicating that there is a direct interaction between the carboxyl-terminal of Cx45 and ZO-1. These studies demonstrate that ZO-1 interacts with Cx45 as well as with Cx43, and suggest that the interaction of connexins with ZO-1 may play a role in regulating the composition of the gap junction and may modulate connexin-connexin interactions.

A Family of 16-kDa Pancreatic Secretory Stress Proteins Form Highly Organized Fibrillar Structures upon Tryptic Activation

A group of 16-kDa proteins, synthesized and secreted by rat pancreatic acinar cells and composed of pancreatic stone protein (PSP/reg) and isoforms of pancreatitis-associated protein (PAP), show structural homologies, including conserved amino acid sequences, cysteine residues, and highly sensitive N-terminal trypsin cleavage sites, as well as conserved functional responses in conditions of pancreatic stress. Trypsin activation of recombinant stress proteins or counterparts contained in rat pancreatic juice (PSP/reg, PAP I and PAP III) resulted in conversion of 16-kDa soluble proteins into 14-kDa soluble isoforms (pancreatic thread protein and pancreatitis-associated thread protein, respectively) that rapidly polymerize into insoluble sedimenting structures. Activated thread proteins show long lived resistance to a wide spectrum of proteases contained in pancreatic juice, including serine proteases and metalloproteinases. In contrast, PAP II, following activation with trypsin or pancreatic juice, does not form insoluble structures and is rapidly digested by pancreatic proteases. Scanning and transmission electron microscopy indicate that activated thread proteins polymerize into highly organized fibrillar structures with helical configurations. Through bundling, branching, and extension processes, these fibrillar structures form dense matrices that span large topological surfaces. These findings suggest that PSP/reg and PAP I and III isoforms consist of a family of highly regulated soluble secretory stress proteins, which, upon trypsin activation, convert into a family of insoluble helical thread proteins. Dense extracellular matrices, composed of helical thread proteins organized into higher ordered matrix structures, may serve physiological functions within luminal compartments in the exocrine pancreas.

Yeast V-ATPase Complexes Containing Different Isoforms of the 100-kDa a-subunit Differ in Coupling Efficiency and in VivoDissociation

The 100 kDa a-subunit of the yeast vacuolar (H(+))-ATPase (V-ATPase) is encoded by two genes, VPH1 and STV1. These genes encode unique isoforms of the a-subunit that have previously been shown to reside in different intracellular compartments in yeast. Vph1p localizes to the central vacuole, whereas Stv1p is present in some other compartment, possibly the Golgi or endosomes. To compare the properties of V-ATPases containing Vph1p or Stv1p, Stv1p was expressed at higher than normal levels in a strain disrupted in both genes, under which conditions V-ATPase complexes containing Stv1p appear in the vacuole. Complexes containing Stv1p showed lower assembly with the peripheral V(1) domain than did complexes containing Vph1p. When corrected for this lower degree of assembly, however, V-ATPase complexes containing Vph1p and Stv1p had similar kinetic properties. Both exhibited a K(m) for ATP of about 250 microm, and both showed resistance to sodium azide and vanadate and sensitivity to nanomolar concentrations of concanamycin A. Stv1p-containing complexes, however, showed a 4-5-fold lower ratio of proton transport to ATP hydrolysis than Vph1p-containing complexes. We also compared the ability of V-ATPase complexes containing Vph1p or Stv1p to undergo in vivo dissociation in response to glucose depletion. Vph1p-containing complexes present in the vacuole showed dissociation in response to glucose depletion, whereas Stv1p-containing complexes present in their normal intracellular location (Golgi/endosomes) did not. Upon overexpression of Stv1p, Stv1p-containing complexes present in the vacuole showed glucose-dependent dissociation. Blocking delivery of Vph1p-containing complexes to the vacuole in vps21Delta and vps27Delta strains caused partial inhibition of glucose-dependent dissociation. These results suggest that dissociation of the V-ATPase complex in vivo is controlled both by the cellular environment and by the 100-kDa a-subunit isoform present in the complex.

Electrophoresis of leaf and seed proteins of Englerophytum natalense (fam. Sapotaceae)

Englerophytum natalense is an indigenous South African tree, bearing edible fruits. In view of its potential for domestication, leaf and seed proteins were separated electrophoretically in order to investigate the existence and degree of variation within and between populations in respect of these proteins. This was part of a process to search for genetic markers, enabling desirable traits to be identified at an early stage in the lives of non-fruiting trees. Acid Polyacrylamide Gel Electrophoresis (A-PAGE) was performed, but yielded no results. Sodium Dodecyl Polyacrylamide Gel Electrophoresis (SDS-PAGE) was successful, both when using leaves of different ages, and when using seeds. A number of variables in the technique were investigated and compared, and optimum parameters are documented. The gels were analysed with the AlphaEaseTMv 3.3 computer package. The densitometric diagrams showed no variation within or between populations; suggesting that either the species is genetically very uniform, or that the proteins investigated do not constitute suitable markers for variation in this species.

Functional and structural properties of the hemoglobin components from Italian sturgeon ( Acipenser naccarii )

The structural and oxygen binding properties of Acipenser naccarii blood have been investigated. The electrophoretic analysis of the hemolysate of this sturgeon showed the presence of two hemoglobin components, each with a considerable globin multiplicity. Constituent globin chains were analyzed by urea-Triton acid polyacrylamide gel electrophoresis and isolated by high performance liquid chromatography. N-terminal amino acid sequence analysis revealed the presence of a N-terminal proline in two of the three α-chains present in the globin pattern, and the presence of a histidine residue in 2βposition. Oxygen equilibria reveal a very low sensitivity of the individual hemoglobins to chloride ions and temperature; however, in the presence of organic phosphates the oxygen affinity of the hemoglobin components decreases strongly. In particular, when Guanosine-5-triphosphate (GTP) is added, the reduction of the oxygen affinity, at pH 7.4 and 20 °C, is 60% and 50%, respectively, for HbI (anodic component) and HbII (cathodic component). As the effect of protons is concerned, the small Root effect shown by total hemolysate at physiological conditions, seems to be due mainly to the cathodic component. On the whole, the functional properties shown by sturgeon hemoglobin components seem to be related to the particular physiological needs dictated by the environmental characteristics.

Abolishment of Proton Pumping and Accumulation in the E1P Conformational State of a Plant Plasma Membrane H+-ATPase by Substitution of a Conserved Aspartyl Residue in Transmembrane Segment 6

The plasma membrane H(+)-ATPase AHA2 of Arabidopsis thaliana, which belongs to the P-type ATPase superfamily of cation-transporting ATPases, pumps protons out of the cell. To investigate the mechanism of ion transport by P-type ATPases we have mutagenized Asp(684), a residue in transmembrane segment M6 of AHA2 that is conserved in Ca(2+)-, Na(+)/K(+)-, H(+)/K(+)-, and H(+)-ATPases and which coordinates Ca(2+) ions in the SERCA1 Ca(2+)-ATPase. We describe the expression, purification, and biochemical analysis of the Asp(684) --> Asn mutant, and provide evidence that Asp(684) in the plasma membrane H(+)-ATPase is required for any coupling between ATP hydrolysis, enzyme conformational changes, and H(+)-transport. Proton pumping by the reconstituted mutant enzyme was completely abolished, whereas ATP was still hydrolyzed. The mutant was insensitive to the inhibitor vanadate, which preferentially binds to P-type ATPases in the E(2) conformation. During catalysis the Asp(684) --> Asn enzyme accumulated a phosphorylated intermediate whose stability was sensitive to addition of ADP. We conclude that the mutant enzyme is locked in the E(1) conformation and is unable to proceed through the E(1)P-E(2)P transition.

Genomic diversity and prevalence of Rotavirus in cow and buffalo calves in northern India.

Faecal samples were collected from seventy-eight diarrhoeic cow and buffalo calves between November 1998 and February 1999 to study the genomic diversity and prevalence of Rotavirus infection by ribonucleic acid polyacrylamide gel electrophoresis (RNA-PAGE) and enzyme-linked immunosorbent assay (ELISA). In the organised dairy farm (where daily production and health records were maintained), the overall prevalence of infection with Rotavirus, recorded by RNA-PAGE and ELISA, was 27.02% (10/37) in both cow and buffalo calves. In unorganised dairy herds (where no production or health records were maintained), RNA-PAGE and ELISA detected infection with Rotavirus in 26.8% (11/41) of cow and 19.5% (8/41) of buffalo calves. Five distinct electropherotypes were found to circulate in cow and buffalo calves. All were short electropherotypes except the single long electropherotype observed in a buffalo calf in an unorganised dairy herd. Some differences in RNA migration pattern were observed when these electropherotypes were compared with the neonatal calf diarrhoea virus strain of Rotavirus. Some electropherotypes were restricted to one farm while others were found in both organised and unorganised dairy herds and in both cow and buffalo calves.

Cleavage of the Actin-capping Protein α-Adducin at Asp-Asp-Ser-Asp633-Ala by Caspase-3 Is Preceded by Its Phosphorylation on Serine 726 in Cisplatin-induced Apoptosis of Renal Epithelial Cells

Decreased phosphorylation of focal adhesion kinase and paxillin is associated with loss of focal adhesions and stress fibers and precedes the onset of apoptosis (van de Water, B., Nagelkerke, J. F., and Stevens, J. L. (1999) J. Biol. Chem. 274, 13328-13337). The cortical actin cytoskeletal network is also lost during apoptosis, yet little is known about the temporal relationship between altered phosphorylation of proteins that are critical in the regulation of this network and their potential cleavage by caspases during apoptosis. Adducins are central in the cortical actin network organization. Cisplatin caused apoptosis of renal proximal tubular epithelial cells, which was associated with the cleavage of alpha-adducin into a 74-kDa fragment; this was blocked by a general caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone (z-VAD-fmk). Hemagglutinin-tagged human alpha-adducin was cleaved into a similar 74-kDa fragment by caspase-3 in vitro but not by caspase-6 or -7. Asp-Arg-Val-Asp(29)-Glu, Asp-Ile-Val-Asp(208)-Arg, and Asp-Asp-Ser-Asp(633)-Ala were identified as the principal caspase-3 cleavage sites; Asp-Asp-Ser-Asp(633)-Ala was key in the formation of the 74-kDa fragment. Cisplatin also caused an increased phosphorylation of alpha-adducin and gamma-adducin in the MARCKS domain that preceded alpha-adducin cleavage and was associated with loss of adducins from adherens junctions; this was not affected by z-VAD-fmk. In conclusion, the data support a model in which increased phosphorylation of alpha-adducin due to cisplatin leads to dissociation from the cytoskeleton, a situation rendered irreversible by caspase-3-mediated cleavage of alpha-adducin at Asp-Asp-Ser-Asp(633)-Ala.

Distinct Signalling Pathways Mediate Insulin and Phorbol Ester-stimulated Eukaryotic Initiation Factor 4F Assembly and Protein Synthesis in HEK 293 Cells

Stimulation of serum-starved human embryonic kidney (HEK) 293 cells with either the phorbol ester, 12-O-tetradecanoylphorbol-13-acetate (TPA), or insulin resulted in increases in the phosphorylation of 4E-BP1 and p70 S6 kinase, eIF4F assembly, and protein synthesis. All these effects were blocked by rapamycin, a specific inhibitor of mTOR. Phosphatidylinositol 3-kinase and protein kinase B were activated by insulin but not by TPA. Therefore TPA can induce eIF4F assembly, protein synthesis, and the phosphorylation of p70 S6 kinase and 4E-BP1 independently of both phosphatidylinositol 3-kinase and protein kinase B. Using two structurally unrelated inhibitors of MEK (PD098059 and U0126), we provide evidence that Erk activation is important in TPA stimulation of eIF4F assembly and the phosphorylation of p70 S6 kinase and 4E-BP1 and that basal MEK activity is important for basal, insulin, and TPA-stimulated protein synthesis. Transient transfection of constitutively active mitogen-activated protein kinase interacting kinase 1 (the eIF4E kinase) indicated that inhibition of protein synthesis and eIF4F assembly by PD098059 is not through inhibition of eIF4E phosphorylation but of other signals emanating from MEK. This report also provides evidence that increased eIF4E phosphorylation alone does not affect the assembly of the eIF4F complex or general protein synthesis.

Electrophoretic identification of hard white spring wheats grown at different locations in Pakistan in different years

Pakistani wheats were identified via acid polyacrylamide gel electrophoresis (A-PAGE) of gliadin proteins. Gliadin electrophoretograms of wheat cultivars were qualitatively independent of growth locations and years. The Pakistani wheat Faisalabad 83 and the American wheat KS84HW196 contained an intense gliadin band in the region of the relative mobility 50 band of Marquis. That band was readily distinguishable and was followed by distinct intense double bands in the same position as those of Marquis, and can be used as a reference for mobility assignments in wheat cultivar identification work. Minor differences in band intensities across locations and years were noticed. Identification tables based on band mobilities relative to Marquis and relative intensities were established.

Studies on Genetic Control of Low-Molecular-Weight Glutenin Subunits in Wheat Endosperm by A-PAGE Analysis

Genetic control of low-molecular-weight glutenin subunits (LMW-GS) of nullisomictetrasomic (NT) and ditelocentric (DT) lines of Chinese Spring, and substitution lines of Cheyenne-Wichiita were investigated using acid-polyacrylamide gel electrophoresis (A-PAGE) analysis. The results showed that genes coding for about 20 LMW-GS could be localized on the short arms (Glu-3 loci) of homologous group 1 chromosomes. On the basis of linkages between Gli-1 loci and Glu-3 loci, several LMW-GS were showed to be coded by different Glu-3 loci by the analysis of F3 generation of Sana x Francuska hybrid. Of 8 LMW glutenin subunits analyzed, 2, 3 and 3 LMW-GS were controlled by Glu-A3, Glu-B3 and Glu-D3 locus, respectively. A-PAGE was capable of rapid, high-resolution separation of glutenin subunits. This system, thus, may become a more efficient alternative to routine SDS-PAGE for investigating composition and genetic control of wheat glutenin subunits.

Floret sterility and outcrossing in two spring wheat cultivars

A tendency for higher outcrossing potential in Canadian semidwarf wheat (Triticum aestivum L.) cultivars compared with tall Canadian cultivars has been postulated by breeders and seed growers. In the present study, the outcrossing potential of a semidwarf Canada Prairie Spring wheat (Triticum aestivum L.) cv. Cutler and a conventional height Canada Western Red Spring wheat, cv. Roblin was determined under controlled greenhouse conditions. Outcrossing of each cultivar was induced by applying moisture stress followed by exposure to pollen from a phenotypic marker stock, cv. P8901. In the controlled absence of external pollen, moisture stress significantly reduced seed set in both cultivars. Application of external pollen following moisture stress increased seed set significantly in Cutler and nonsignificantly in Roblin, and was associated with a higher level of floret opening in Cutler. Outcrossing frequency was also assessed by analysing progeny seed (selfed or outcrossed) using acidic polyacrylamide gel electrophoresis of seed protein, RAPD markers and morphological phenotype, including height, awnedness, black chaff and time to maturity. All three assay techniques demonstrated a higher outcrossing frequency in Cutler than in Roblin under this protocol. The frequency of outcrossing in different parts of the spike was also determined for both cultivars. In both cultivars the highest proportion of outcrossing was found in the mid-upper region of the spike, followed by the mid-lower region. Key words: Floret sterility, moisture stress, outcrossing, Triticum aestivum

Acid-polyacrylamide gel electrophoresis of lysozyme-estrone glucuronide conjugates.

Acid-polyacrylamide gel electrophoresis (acid-PAGE) was used for analysis of lysozyme-estrone glucuronide conjugates. The resolution of the system allowed the identification of individual conjugate families which differed only in the position of acylation or in the number of estrone glucuronide units. Acid-PAGE was a good alternative to denaturing cation-exchange chromatography for the analysis, separation, and small-scale purification of lysozyme-estrone glucuronide conjugates. It revealed the true order of the relative degree of positive charge on the lysozyme-estrone glucuronide conjugates.

Characterization of the Interactions of Plasminogen and Tissue and Vampire Bat Plasminogen Activators with Fibrinogen, Fibrin, and the Complex of d-Dimer Noncovalently Linked to Fragment E

Vampire bat plasminogen activator (b-PA) causes less fibrinogen (Fg) consumption than tissue-type plasminogen activator (t-PA). Herein, we demonstrate that this occurs because the complex of D-dimer noncovalently linked to fragment E ((DD)E), the most abundant degradation product of cross-linked fibrin, as well as Fg, stimulate plasminogen (Pg) activation by t-PA more than b-PA. To explain these findings, we characterized the interactions of t-PA, b-PA, Lys-Pg, and Glu-Pg with Fg and (DD)E using right angle light scattering spectroscopy. In addition, interactions with fibrin were determined by clotting Fg in the presence of various amounts of t-PA, b-PA, Lys-Pg, or Glu-Pg and quantifying unbound material in the supernatant after centrifugation. Glu-Pg and Lys-Pg bind fibrin with Kd values of 13 and 0.13 microM, respectively. t-PA binds fibrin through two classes of sites with Kd values of 0.05 and 2.6 microM, respectively. The second kringle (K2) of t-PA mediates the low affinity binding that is eliminated with epsilon-amino-n-caproic acid. In contrast, b-PA binds fibrin through a single kringle-independent site with a Kd of 0.15 microM. t-PA competes with b-PA for fibrin binding, indicating that both activators share the same finger-dependent site on fibrin. Glu-Pg binds (DD)E with a Kd of 5.4 microM. Lys-Pg binds to (DD)E and Fg with Kd values of 0.03 and 0.23 microM, respectively. t-PA binds to (DD)E and Fg with Kd values of 0.02 and 0.76 microM, respectively; interactions were eliminated with epsilon-amino-n-caproic acid, consistent with K2-dependent binding. Because it lacks a K2-domain, b-PA does not bind to either (DD)E or Fg, thereby explaining why b-PA is more fibrin-specific than t-PA.

Effects of wheat bug (Eurygaster maura) proteolytic enzymes on electrophoretic properties of gluten proteins

Hydrolysis of gluten proteins of six bread wheat (Triticum aestivum L.) cultivars damaged by Eurygaster maura was investigated by acid polyacrylamide gel electrophoresis (A‐PAGE) and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS‐PAGE). Electrophoresis results showed that bug proteolytic enzymes clearly affected gliadin and glutenin proteins of damaged wheats. Some new bands of low and high mobility appeared in the A‐PAGE electrophoregrams. These new bands and the bands in the original gliadin and glutenin patterns disappeared with increasing incubation times. Especially high molecular weight (HMW) glutenin subunits were most strongly affected by the bug protease, and disappeared at 120 min of incubation (except ‘Ankara'). However, some of the low molecular weight (LMW) glutenin subunits were not affected as much as HMW glutenin subunits and were still visible even after the longest incubation time investigated. Electrophoretic patterns of some cultivars with the same level of proteolytic activity were affected differently, indicating an intercultivar variation in susceptibility to hydrolysis by bug proteolytic enzymes.

Activity staining of pectinesterase on polyacrylamide gels after acidic or sodium dodecyl sulfate electrophoresis.

Pectinesterase (PE), from commercial orange peels or ammonium sulfate fractionation (50-80% saturation) of pea pods, was detected on polyacrylamide gels after native acidic polyacrylamide gel electrophoresis (PAGE) or sodium dodecyl sulfate (SDS)-PAGE by using the synthetic substrate beta-naphthyl acetate (beta-NA). The release of beta-naphthol (at 322 nm) from beta-NA was proportional to PE activity. The PE activity bands on polyacrylamide gels after native acidic PAGE or SDS-PAGE were stained with a combination of tetrazotized o-dianisidine and beta-NA. This fast and sensitive method can be used for enzyme purification and characterization.

Characterisation of European oat germ plasm: allelic variation at complex avenin loci detected by acid polyacrylamide gel electrophoresis

European oat germ plasm involving 252 varieties, landraces, and breeding lines has been characterised by allelic constitution at three independent complex avenin loci. A total of 26 avenin alleles (blocks), including 7 at the Ave1 (Avn A), 10 at the Ave2 (Avn B), and 9 at the Ave3 (Avn C) loci were revealed. The avenin block variants were shown to be coded by gene clusters of 2–5 single expressed genes. A catalogue of the identified avenin blocks as well as a list of the avenin phenotypes is presented. It has been discovered that about 8% of the oat varieties were heterogeneous comprising two or three different avenin profiles. The varieties assayed have been classified into 79 avenin phenotypes made up by the different alleles. Forty-six of these phenotypes were distinguished uniquely, while the other 33 were shared by groups possessing from 2 to 41 genotypes each. Members within these groups had certain ancestors in common. Non-random patterns of both the avenin allele and phenotype distributions have been found.

Gel electrophoretic investigations of prolamins in eu- and alloplasmatic octoploid primary triticale forms

Unreduced prolamin fractions of eu- and alloplasmatic octoploid triticale forms were investigated by means of gel electrophoresis. The electrophoretic separation of the prolamin fractions was carried out by using one-dimensional, horizontal, native acidic polyacrylamide gel electrophoresis (APAGE) and gradient gels. A comparison of the electropherogram patterns of triticale forms with those of the genome donors showed that most of the parent prolamins can be found again in the corresponding triticale. However, the bands of the triticale forms exhibited lower intensities than those of the genome donors. On average the gliadin bands of the triticale forms showed equal reductions in intensities, thereby preserving the relationships of the band intensities in most cases. On the other hand, secalin bands which came from the rye parent showed varying reductions in intensities. A comparative analysis of the band patterns revealed that differences could often be found between triticale forms with the same genomic constitution. Often it concerned differences in intensity. In some cases the bands showed changes in mobility or a splitting up into two sub-bands. Starting with the plant material presented here we developed a new nomenclature of the prolamin band patterns of the triticale forms.