Major polypeptides associated with differentiation in psychrophilic fungi

1987 ◽  
Vol 65 (2) ◽  
pp. 233-241 ◽  
Author(s):  
W. J. Newsted ◽  
N. P. A. Huner

Major polypeptides were observed upon one-dimensional sodium dodecyl sulfate gel electrophoresis of sclerotial extracts of the following psychrophiles: Myriosclerotinia borealis, Coprinus psychromorbidus, Typhula idahoensis, and Typhula incarnata. In general, the number, molecular mass, and relative proportion of these major sclerotial polypeptides varied considerably from species to species. Furthermore, in the case of M. borealis, the major sclerotial polypeptide did not appear to be an artifact of culturing conditions since a major polypeptide of similar molecular mass was also present in sclerotia of M. borealis collected from the field. Generally, the major sclerotial polypeptides were visible in the sclerotial initials but were not apparent in the vegetative hyphae. Thus, these major sclerotial polypeptides appear to be expressed as a function of sclerotial development. Electrophoresis of protein extracts of T. idahoensis and T. incarnata initially solubilized either in sodium dodecyl sulfate or urea sample buffer indicated that the type of denaturant initially used had a profound influence on the relative proportions of the major polypeptides and the overall polypeptide profile. Isoelectric focusing of sclerotial extracts indicated that the isoelectric points of the major sclerotial polypeptides of M. borealis ranged from 6.2 to 6.7, whereas the values of the major sclerotial polypeptides of the other three species were basic and ranged from 7.0 to 7.7.

1985 ◽  
Vol 63 (12) ◽  
pp. 2311-2318 ◽  
Author(s):  
W. J. Newsted ◽  
N. P. A. Huner ◽  
J. P. Insell ◽  
M. Griffith ◽  
R. B. van Huystee

Myriosclerotinia borealis (W51), Coprinus spp. (13W1 and 14W2), Typhula idahoensis (W21), and Typhula incarnata (W29) were incubated in the dark on a defined agar medium at permissive (4 °C) and nonpermissive temperatures (22 and 30 °C). Isolates of Coprinus spp. and Typhula spp. required higher temperatures than M. borealis to arrest vegetative growth completely. The effects of incubation at permissive and nonpermissive temperatures on the polypeptide compositions of M. borealis, Coprinus spp., T. idahoensis (W21), and T. incarnata (W29) were examined by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The results indicated that the number of polypeptides in the polypeptide complement of M. borealis and both Typhula species decreased significantly during incubation at nonpermissive temperatures. In contrast, Coprinus sp. (13W1) showed no significant change in the number of polypeptides observed during incubation at nonpermissive temperatures. Furthermore, there appeared to be an increase in the relative proportion of at least three polypeptides during incubation of Coprinus at nonpermissive temperatures. The significance of these species-dependent responses to nonpermissive growth temperatures is discussed.


1986 ◽  
Vol 32 (5) ◽  
pp. 811-815 ◽  
Author(s):  
J M Perini ◽  
B Dehon ◽  
T Marianne ◽  
A Klein ◽  
P Roussel

Abstract Immunoblots of several urinary low-molecular-mass proteins can be very useful in investigations of pathological proteinuria. However, use of certain commercial antisera in such procedures leads to artifacts corresponding to nonspecific bands; e.g., immunoglobulins from nonimmunized rabbit serum may bind to human urinary proteins, and this binding is not inhibited by Triton X-100. We have developed a procedure to improve the specificity of detection of urinary low-Mr proteins separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, by immunoblotting with commercial antisera: we treat the protein blot with a mixture of mercaptoethanol and sodium dodecyl sulfate before incubation with the first antiserum. This allows direct use of commercial antisera without prior absorption of contaminating antibodies.


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