Electrophoretic profiles of cyanobacterial membrane polypeptides showing heme-dependent peroxidase activity

The photosynthetic membranes of Anacystis nidulans R2 were examined electrophoretically following solubilization with lithium dodecyl sulfate. Electrophoresis yielded six prominent chlorophyll-containing bands. In addition, five polypeptides were observed which possessed heme-dependent peroxidase activity, monitored by incubating gels with 3,3′,5,5′-tetramethylbenzidine plus hydrogen peroxide. One such polypeptide, at 105 kdaltons, was removed by repeated washing of the membranes. Four remaining peroxidase-active polypeptides were observed at 7.2, 13.5, 18.5 and 33 kdaltons. Further examination of these four polypeptides yielded the following results. (1) The mobility of the 33 kdalton polypeptide was altered from 29 to 33 kdaltons upon heating (70°C) during membrane solubilization. (2) All four polypeptides showed stable heme-protein associations in the presence of 8 M urea; however, in the presence of urea, alterations in protein mobility were observed for each poly-peptide and only two (at 13.5 and 33 kdaltons) showed peroxidase activity following heating (70°C) during membrane solubilization. (3) The presence of thiols during membrane solubilization at 0°C was required to observe peroxidase activity at 7.2 kdaltons. These results, when compared to known properties of isolated cytochromes, suggest that the four polypeptides characterized here correspond to the subunits of photosynthetic cytochromes. Electrophoretic assessment of maize mutants lacking cytochrome f and b₆ activity supports this suggestion.     

Characterization of the DNA-cellulose-binding proteins from Escherichia coli K 12

The various [35S]DNA-binding proteins present in lysates of Escherichia coli K 12 cells have been analyzed by means of two-dimensional SDS-polyacrylamide gel electrophoresis. The proteins were isolated by the DNA-cellulose technique and eluted by increasing concentrations of NaCl (0.15, 0.4, 0.6 and 2 M). Only 2% of the total 35S radioactivity in the lysate became bound to the DNA-cellulose column. A total of 237 polypeptides were detected and the distribution among the salt eluates were 85, 83, 40 and 29 polypeptides, respectively. The 40 major polypeptides with regard to concentrations were also identified from gels stained with a protein-specific reagent. The polypeptides could be divided into two main groups according to pI values, namely, acidic polypeptides (total number, 174) and basic polypeptides (total number, 63). The ratio between acidic and basic polypeptides decreased with increasing salt concentrations in the eluates. The majority of the basic polypeptides had molecular weights in the range 10 000-30 000, whereas the acidic polypeptides had molecular weights from 10 000 to 165 000.     

Modification of lipophilic proteins in Friend erythroleukemia cells during their differenciation

In mouse erythroleukemia cells (MELC) a lipophilic protein of apparent M. W. 9.5 kdaltons increases during differentiation. This increase is due either to an increase of biosynthesis or to a structural alteration impairing the capacity of the protein to form polymers of apparent high M.W. or favoring its extractability. The increase is related to differentiation and precedes hemoglobin synthesis by at least 1 day. It is not related cells (TFA-11) in which dimethylsulfoxide (DMSO) causes an increase in virus production. As it occurs in cells treated with 4 different inducers, and as the increase is less marked when antagonists of the inducers are also present, it is unlikely that the increase of the 9.5 Kdalton protein is due to an effect of the inducers unrelated to differentiation.     

Conformational and aggregation properties of a PEGylated alanine-rich polypeptide

The conformational and aggregation behavior of PEG conjugates of an alanine-rich polypeptide (PEG-c17H6) were investigated and compared to that of the polypeptide equipped with a deca-histidine tag (17H6). These polypeptides serve as simple and stimuli-responsive models for the aggregation behavior of helix-rich proteins, as our previous studies have shown that the helical 17H6 self-associates at acidic pH and converts to β-sheet structures at elevated temperature under acidic conditions. In the work here, we show that PEG-c17H6 also adopts a helical structure at ambient/subambient temperatures, at both neutral and acidic pH. The thermal denaturation behavior of 17H6 and PEG-c17H6 is similar at neutral pH, where the alanine-rich domain has no self-association tendency. At acidic pH and elevated temperature, however, PEGylation slows β-sheet formation of c17H6, and reduces the apparent cooperativity of thermally induced unfolding. Transmission electron microscopy of PEG-c17H6 conjugates incubated at elevated temperatures showed fibrils with widths of ～20-30 nm, wider than those observed for fibrils of 17H6. These results suggest that PEGylation reduces β-sheet aggregation in these polypeptides by interfering, only after unfolding of the native helical structure, with interprotein conformational changes needed to form β-sheet aggregates.     

Acidic ribosomal proteins of Neurospora crassa

Neurospora crassa acidic ribosomal proteins from the high salt-ethanol extract of 80 S ribosomes have been fractionated by DEAE-cellulose chromatography. Six acidic ribosomal proteins were purified. All resemble Escherichia coli L7 and L12 in amino acid composition and molecular weight but each has a slightly different net charge at pH 3.2. Four have an apparent molecular weight of approx. 14 000, and two have a molecular weight of approx. 14 800. The amino acid compositions and circular dichroism (CD) spectra of the purified Neuropsora proteins are identical for the four 14 kDa proteins, but clearly distinguishable from the two 14.8 kDa proteins. The latter are also identical in amino acid composition and CD spectra. This suggests that there are two Neurospora acidic, or 'A', proteins, one of which exists in four microheterogeneous forms and the other exists in two forms.     

Salt-induced changes in protein composition in light-grown callus of Mesembryanthemum crystallinum

The halophyte Mesembryanthemum crystallinum (ice plant) has been suggested as a model for salt-tolerance in higher plants. To investigate salt-induced changes in polypeptide patterns at the cellular level, a light-grown callus of M. crystallinum with substantial chlorophyll content, was established and the effect of NaCl on the composition of phenol-extracted protein was examined by SDS- and 2D-polyacrylamide gel electrophoresis (PAGE). SDS-PAGE showed the accumulation of five polypeptides with estimated molecular masses of 40, 34, 32, 29 and 14 kDa was enhanced by the addition of 200 m M NaCl to the culture media. The addition of ABA (10 μ M ) or mannitol (400 m M ) did not elicit the same degree of accumulation of these salt-specific proteins. These polypeptides were classified into two groups according to their course of induction: early responsive (40, 34, 29 kDa) and late-responsive (32, 14 kDa) proteins. In addition, two polypeptides (20, 18 kDa) were transiently accumulated during salt treatment. Further separation of soluble proteins by 2-D gel electrophoresis, either isoelectric focusing (IEF) or non-equilibrium pH-gradient electrophoresis (NEPHGE) followed by SDS-PAGE, showed more alterations in accumulation of polypeptides by NaCl than 1-D gel electrophoresis. Overall, levels of more than 30% of basic polypeptides, detected by NEPHGE/SDS-PAGE, were altered by 200 m M NaCl treatment, while only 10% of neutral and acidic polypeptides, detected by IEF/SDS-PAGE, were changed. The enhanced expression of these proteins by salt in cultured cells is most likely related to the cellular responses to salinity, and not to the mechanism of CAM induction in this facultative halophyte.     

Surface plasmon resonance characterization of calspermin-calmodulin binding kinetics

We cloned, expressed, and purified a chimeric fusion between a soluble green fluorescent protein (smGFP) and the calmodulin binding protein calspermin. We have shown that the fusion protein, labeled smGN, has a K(i) in the calmodulin-dependent cyclic nucleotide phosphodiesterase activity assay of 1.97 nM, i.e., 3800 times smaller than that of the commonly used calmodulin inhibitor W7. Association and dissociation rate constants (k(a) and k(d)) and the dissociation equilibrium constant (K(D)) of smGN for calmodulin were determined using surface plasmon resonance (SPR). The k(a)=1.24 x 10(6)M(-1)s(-1), the k(d)=5.49 x 10(-3)s(-1), and the K(D)=4.42 x 10(-9)M. We also found that the GFP moiety was important for successfully binding calspermin to the surface of the CM5 flow cell at a sufficiently high concentration for SPR, and that this procedure may be used for SPR analysis of other acidic polypeptides, whose pI< or =4. To determine whether smGN might also bind to other calmodulin-like proteins in a heterologous system, we purified proteins from a plant total cell extract or a plant total protein extract by affinity chromatography against smGN. The purified proteins were identified as calmodulins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and liquid chromatography-tandem mass spectrometry, indicating a high level of specificity. We conclude that the high affinity and specific binding between smGN and calmodulin make it an easily localized recombinant alternative to chemical calmodulin inhibitors.     

Synthesis and incorporation of myelin polypeptides into CNS myelin

The distribution of newly synthesized proteolipid protein (PLP, 23 kdaltons) and myelin basic proteins (MBPs, 14-21.5 kdaltons) was determined in microsomal and myelin fractions prepared from the brainstems o1 10-30 d-old rats sacrificed at different times after an intracranial injection of 35S-methionine. Labeled MBPs were found in the myelin fraction 2 min after the injection, whereas PLP appeared first in the rough microsomal fraction and only after a lag of 30 min in the myelin fraction. Cell-free translation experiments using purified mRNAs demonstrated that PLP and MBPs are synthesized in bound and free polysomes, respectively. A mechanism involving the cotranslational insertion into the ER membrane and subsequent passage of the polypeptides through the Golgi apparatus is consistent with the lag observed in the appearance of the in vivo-labeled PLP in the myelin membrane. Newly synthesized PLP and MBPs are not proteolytically processed, because the primary translation products synthesized in vitro had the same electrophoretic mobility and N-terminal amino acid sequence as the mature PLP and MBP polypeptides. It was found that crude myelin fractions are highly enriched in mRNAs coding for the MBPs but not in mRNA coding for PLP. This suggests that whereas the bound polysomes synthesizing PLP are largely confined to the cell body, free polysomes synthesizing MBPs are concentrated in oligodendrocyte processes involved in myelination, which explains the immediate incorporation of MBPs into the developing myelin sheath.     

Protein synthesis and transport in the regenerating goldfish visual system

The nature of the proteins synthesized in the goldfish retina and axonally transported to the tectum during optic nerve regeneration has been examined. Electrophoretic analysis of labeled soluble retinal proteins by fluorography verified our previous observation of a greatly enhanced synthesis of the microtubule subunits. In addition, labeling of a tubulin-like protein in the retinal particulate fraction was also increased during regeneration. Like soluble tubulin, the particulate material had an apparent MW of 53–55K and could be tyrosylated in the presence of cycloheximide and [³H]tyrosine. Comparison of post-crush and normal retinal proteins by two-dimensional gel electrophoresis also revealed a marked enhancement in the labeling of two acidic 68–70K proteins. Analysis of proteins slowly transported to the optic tectum revealed changes following nerve crush similar to those observed in the retina, with enhanced labeling of both soluble and particulate tubulin and of 68–70K polypeptides. The most striking change in the profile of rapidly transported protein was the appearance of a labeled 45K protein which was barely detectable in control fish.     

The 55-kDa polypeptide released from spinach thylakoid membranes with 1 M LiCl is not the beta subunit of chloroplast F1

It was reported by Frasch et al. (Frasch, W. D., Green, J., Caguiat, J., and Mejia, A. (1989) J. Biol. Chem. 264, 5064-5069) that washing spinach thylakoid membranes with 1 M LiCl caused the release of the beta subunit of chloroplast F1 (CF1) which, existing as 180-kDa complexes of beta 3, retained considerable ATPase activity. We repeated their procedures and confirmed that a CF1 beta-like 55-kDa polypeptide was a major constituent of the 1 M LiCl-washed extract. However, the extract contained another polypeptide of which the Mr was 14,000, and these two polypeptides comprised a complex with approximate Mr 550,000 that had the same mobility in native polyacrylamide gel electrophoresis as that of ribulose-1,5-bisphosphate carboxylase. Only very low ATPase activity, less than 1% of the reported value, was detected for the extract and the purified complex. Antibody against the beta subunit of F1 from a thermophilic bacterium PS3 showed a clear cross-reactivity with the CF1 beta subunit but not with the 55-kDa polypeptide. Analysis of the N-terminal amino acid sequences of the 55- and 14-kDa polypeptides and the whole complex revealed that the complex was ribulose-1,5-bisphosphate carboxylase and that the 55- and 14-kDa polypeptides were its large and small subunits, respectively.     

水稻谷蛋白是类似于豆球蛋白的蛋白质

水稻是我国的主要粮食作物,它的蛋白质含量为5—14%。在水稻种子蛋白质中,谷蛋白占80%,球蛋白占10%,醇溶蛋白占5%,清蛋白占5%。据报道,水稻种子清蛋白主要由分子量16800的亚基组成,球蛋白由10种不同分子量的亚基通过疏水交互作用相结合,谷蛋白由分子量38000、25000和16000三种亚基通过双硫键相结合:但是,Yamagata(1982)和作者(1983、1984、1986)的研究表明:水稻种子谷蛋白主要由分子量     

Multivariate analysis of polypeptide synthesis in field-grown maize inbreds and hybrids

Summary A leaf disc method is described to permit the localized incorporation of ³⁵S-l-methionine into polypeptides synthesized in individual leaves of maize plants grown in the field. The method of incorporation employs minimal external manipulation of the intact leaf, is simple, repeatable, and may be used at any plant age after leaf emergence. Incorporation (cpm/g protein) in 12 leaves per plant was compared among three inbred (Oh43, W23, M14) and three F₁ hybrid (Oh43/M14, W23/M14, Oh43/W23) genotypes. The incorporation was 40% higher (hybrid versus inbred) in 9 of the 12 leaves studied. Samples from leaf 07 (7th leaf numbered from base of plant) for four inbreds (Oh43, M14, B73, Mol 7) and two pairs of reciprocal F₁ hybrids (Oh43/M14, M14/Oh43; B73/Mo17, Mo17/B73) were labelled in situ using the leaf disc method. Each cultivar was sampled at three different ages in each of 1985, 1986, and 1987. High-resolution, two-dimensional isoelectric focusing sodium-dodecyl-sulfate polyacrylamide gel electrophoresis and fluorography were used to display the polypeptides synthesized in the samples. Multivariate methods — Principal Coordinate Analysis, Cluster Analysis, and Standard Deviation Distance — were used to analyze variation and to identify trends in the variation for year, genotype, and age sampled. Our analyses disclose a hierarchy to polypeptide synthesis variation in maize leaves: differences in polypeptide synthesis are greater for year-to-year comparisons than differences due to sample age, which in turn are greater than differences for inbred versus hybrid comparisons.     

Calmodulin and acidic compounds alter basic protein phosphorylation by the protein kinase from human platelets

A cyclic nucleotide-independent protein kinase of human platelets, which phosphorylated histones, myelin basic protein and protamine and did not catalyze the phosphorylation of acidic proteins such as casein, phosvitin and myosin light chain, has been purified approx. 1,500-fold from the crude extract by steps of DEAE-cellulose, Sephadex G-200, hydroxylapatite and phosphoryl cellulose column chromatography. The substrate phosphorylation by this kinase was markedly enhanced by calmodulin even in the absence of Ca²⁺, when mixed histone was used as a substrate. The interaction of the kinase with mixed histone resulted in an irreversible inactivation of the enzyme. Calmodulin prevented this inactivation, and this compound produced an apparent increase in histone phosphorylation by the kinase. It should be noted that acidic polypeptides such as troponin-C, phospholipids and nucleic acids have a similar ability. The addition of Ca²⁺ reduced the effect of calmodulin more than the effects of other acidic compounds.     

Specific, water-soluble polypeptides in identified neurons of Aplysia californica.

Application of an ethylene glycol lysis technique to extract water-soluble, low molecular weight polypeptides in Aplysia neurons, was used in conjunction with microgradient gel electrophoresis and micro-isoelectric focusing, to identify unique polypeptides in specific, identified neurons. The polypeptides found in neurons R15, R3-13, R14, and the bag cells were particularly abundant, consistent with the previously suggested neurosecretory role for these cells. Water extraction of the strongly basic polypeptides (pI 10.7) in R3-13 and R14 required an acidic lysis medium.     

Isolation and identification of antimicrobial components from the epidermal mucus of Atlantic cod (Gadus morhua)

The epidermal mucus of fish species has been found to contain antimicrobial proteins and peptides, which is of interest in regard to fish immunity. An acidic extract from the epidermal mucus of the Atlantic cod (Gadus morhua) was found to exhibit antimicrobial activity against Bacillus megaterium, Escherichia coli and Candida albicans. This activity varied significantly when salt was added to the antimicrobial assay, and was eliminated by pepsin digestion. No lysozyme activity was detected in the extract. By using weak cationic exchange chromatography together with reversed-phase chromatography, and monitoring the antimicrobial activity, we have isolated four cationic proteins from the mucus extract. Using N-terminal and C-terminal amino acid sequence analysis, together with MS, the antimicrobial proteins were identified as histone H2B (13 565 Da), ribosomal protein L40 (6397 Da), ribosomal protein L36A (12 340 Da) and ribosomal protein L35 (14 215 Da). The broad spectra of antimicrobial activities in the cod mucus and the characterization of four antimicrobial polypeptides suggest that mucus compounds contribute to the innate host defence of cod.     

Effect of fungal-isolate aggressivity on the biosynthesis of symbiosis-related polypeptides in differentiating eucalypt ectomycorrhizas

Changes in protein biosynthesis were examined during the early stages of differentiation of Eucalyptus grandis-Pisolithus tinctorius ectomycorrhizas by two-dimensional polyacrylamide gel electrophoresis of ³⁵S-labelled proteins. Three distinct isolates of P. tinctorius Coker & Couch were chosen based on the rate of ectomycorrhizal formation (i.e. infectivity) with E. grandis W. Hill ex Maiden. The isolate H506 was not able to induce mycorrhiza, isolate 441 showed moderate infectivity and isolate H2144 exhibited a very high infectivity. Mycorrhiza were produced in vitro in a system where seeds were germinated in the presence of fungal mycelium and exudates. The non-mycorrhizal isolate caused no changes in root protein biosynthesis as analyzed by two-dimensional polyacrylamide gel electrophoresis, whereas drastic alterations in protein biosynthesis were observed from initial contact with the aggressive mycobionts. During mycorrhizal development, there was a marked inhibition of plant polypeptides synthesis, enhanced accumulation of some fungal polypeptides and the emergence of symbiosis-specific polypeptides, the so-called ectomycorrhizins. The major changes were observed in a group of fungal acidic polypeptides (apparent molecular weight 28–32 kDa) including the ectomycorrhizin E₃₂. These polypeptides first appeared at contact and their synthesis increased during mycorrhizal formation, suggesting a role in mycorrhizal development, most likely as structural proteins. Up-regulation of the synthesis of fungal symbiosis-related polypeptides was tightly correlated to the infectivity of the strain.Abbreviations FW fresh weight - MW molecular weight - pI isoelectric point - SR-polypeptides symbiosis-related polypeptides This work was supported by a research grant from the Eureka-Eurosilva programme (Changes in Gene Expression during Ectomycorrhiza Differentiation and Function) to F.M. and a Murdoch University Special Research Grant to B.D; T.B. was a recipient of a Doctoral Fellowship from the INRA and an Australian Postgraduate Scholarship. We would like to thank Dr Denis Tagu and Dulcinéia de Carvalho (Institut National de la Recherche Agronomique, Nancy, France) for helpful discussions.     

Effect of heat and 2-mercaptoethanol on intracytoplasmic membrane polypeptides of Rhodopseudomonas sphaeroides.

Solubilization at 75 degrees C of Rhodopseudomonas sphaeroides chromatophores in the presence of sodium dodecyl sulfate (SDS) and 2-mercaptoethanol (beta-ME) resulted in the selective absence of reaction center B and C polypeptides from SDS-polyacrylamide gel electrophoresis profiles. A newly identified, chromatophore-specific polypeptide, with a mass of 35.2 kdaltons, was also missing under these conditions of chromatophore solubilization. Solubilization at 27 degrees C in the presence of SDS and beta-ME also resulted in the disappearance of these three polypeptides, but at much slower rates. Disappearance of either endogenous or exogenously supplied reaction center polypeptides B and C during SDS solubilization of whole chromatophores at either 27 or 75 degrees C was shown to be entirely dependent upon the presence of beta-ME. After chromatophore solubilization in the presence of beta-ME and subsequent SDS-polyacrylamide gel electrophoresis, exogenously added reaction centers B and C could be localized in a complex of no less than 100 to 200 kdaltons. However, the precise size of the complex was influenced by the stoichiometry of the reacting components. The disappearance of the 35.2-kdalton polypeptide was neither dependent upon the presence of beta-ME nor dependent upon the presence of any additional chromatophore polypeptides. The 35.2-kdalton polypeptide underwent a heat-induced oligomerization to yield several high-molecular-weight species.     

Variations in Brain Cortical Polysome Translation Products Among Rats of the Same Strain

Studies were undertaken to determine whether there exist variations among the translation products of polysomes from different brains of animals of the same strain. Polysomes were prepared from individual rat cortices and translated in a reticulocyte protein-synthesizing system containing rabbit reticulocyte factors and L-[35S]methionine; he resulting radioactive proteins were analyzed by two-dimensional polyacrylamide gel electrophoresis autoradiography. Comparison of the autoradiographs revealed that two acidic proteins, A and B, of apparent 54,000 M. W. occur as three phenotype: A only, B only, or A plus B. These proteins were not detectable by Coomassie brilliant blue staining of two-dimensional electrophoretograms of brain protein preparations. Messenger RNA was extracted from pooled cortices and translated in a wheat germ extract, and both A and B proteins were detected among the products of translation. Cyclic AMP affinity chromatography of the translation products of cortical polysomes showed that both A and B proteins bind to cyclic AMP. Our data are consistent with the conclusion that there are qualitative differences in the polysome translation products that bind to cyclic AMP among individual cortices of rats of the same strain.     

Organization of Brain Synaptic Vesicle Proteins

Abstract: The topographical arrangement of proteins and glycoproteins of mouse brain synaptic vesicles was studied with trypsin and galactose oxidase, reagents known to be impermeable with respect to other membranes. Incubation of vesicles with trypsin at a concentration of 1 μg/ml extensively degraded seven polypeptides of molecular weights (M.W.) (×10^-3) 125, 107, 95, 83, 70, 60, and 36; higher concentrations degraded two additional species of 75,000 and 46,000 M.W., while leaving unaffected polypeptides of M.W. 66,000, 55,000, 33,000, 26,000, 22,000, 19,000, and 16,000. All of the trypsin-sensitive species of greater than 70,000 M.W. stained positively with the periodic acid-Schiff reagent; several other glycoproteins, all of M.W. less than 70,000, were identified, and all of these were insensitive to trypsin. Galactose oxidase-NaB³H₄ treatment of synaptic vesicles heavily and exclusively labeled material of greater than 70,000 M.W. All of the polypeptides studied were sensitive to each reagent when the synaptic vesicles were first treated with detergents. Extraction of vesicles with 0.05 M-NaOH partially or completely removed a wide variety of polypeptides, including most of those in the M.W. range 46,000–83,000; none of the glycoproteins was solubilized. Essentially the opposite results were obtained when the vesicles were extracted with 0.5% Triton X-100. Most of the vesicle's species were insensitive to several bisimidate cross-linking reagents. These results suggest that: (a) The polypeptides of M.W. 125K, 107K, 95K, 83K, 75K, 70K, 60K, 46K, and 36K are externally oriented in the vesicle, whereas those of 66K, 55K, 33K, 26K, 22K, 19K, and 16K are internally oriented; (b) the vesicles contain two classes of glycoproteins, one consisting of high-molecular-weight, externally oriented species that are rich in galactose, and the other consisting of low-molecular-weight, internally oriented species of relatively low galactose content; (c) the vesicles contain a large class of nonglycosylated species that are relatively loosely attached to the membrane; and (d) most of the vesicles' polypeptides are probably freely mobile in the membrane. The organization of synaptic vesicle proteins is compared with that of the proteins of synaptosomal plasma membrane, with which the vesicle is believed to fuse.     

Major sclerotial polypeptides of psychrophilic pathogenic fungi: Intracellular localization and antigenic relatedness

Summary Major sclerotial polypeptides from the psychrophiles,Myriosclerotinia borealis (W 51),Coprinus psychromorbidus (LRS 131),Typhula idahoensis (W 21), andTyphula incarnata (W 21) were purified by using polyacrylamide gel electrophoresis and electroelution. Polyclonal antibodies were raised against these major sclerotial polypeptides. Immunofluorescence microscopy showed that the major sclerotial polypeptides from all four psychrophilic species were sequestered in discrete protein bodies of cultured and field-grown sclerotia. Western blot analysis indicated that all antisera reacted positively with their respective antigens, the major sclerotial polypeptides. Reciprocal immunological cross-reactions were observed between the major sclerotial polypeptides ofM. borealis (W 51) andT. idahoensis (W 21). Antiserum to the major sclerotial polypeptides of bothM. borealis andT. idahoensis also recognized the major sclerotial polypeptides ofC. psychromorbidus (LRS 131). It is suggested that the major sclerotial polypeptides of these psychrophilic plant pathogens may act as storage proteins.Abbreviations W 51 Myriosclerotinia borealis (W 51) - LRS 131 Coprinus psychromorbidus (LRS 131) - W 21 Typhula idahoensis (W 21) - W 29 Typhula incarnata (W 29) - anti W 51 antiserum to the major sclerotial polypeptide ofM. borealis W 51 - anti LRS 131 antiserum to the major sclerotial polypeptides ofC. psychromorbidus (LRS 131) - anti W 21 antiserum to the major sclerotial polypeptides ofT. idahoensis (W 21) - anti W 29 antiserum to the major sclerotial polypeptides ofT. incarnata (W 29) - SDS sodium dodecylsulfate - kDa kilodalton - PAGE polyacrylamide gel electrophoresis - HRP horseradish peroxidase - PBS phosphate buffered saline - TBS Tris buffered saline - FITC fluorescein isothiocyanate