Immunochemistry and localization of the enzyme disaggregatase in Methanosarcina mazei.
The enzyme disaggregatase (Dag) from Methanosarcina mazei was studied immunochemically. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of purified Dag under reducing and nonreducing conditions revealed a single band with a 94-kDa molecular mass. Dag was found to be immunogenic in rabbits; a polyclonal antibody probe was prepared and used to detect the enzyme by slide immunoenzymatic assay, immunofluorescence, and immunoblotting in various species of Methanosarcina known to convert from packets to single cells, including M. mazei. The enzyme could not be detected in other members of the family Methanosarcinaceae that do not convert. By immunogold electron microscopy, Dag was mapped to the cell wall of packets and to the cell membrane of single cells of two M. mazei strains.
Conway de Macario
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E
Wadsworth Center for Laboratories and Research, New York State Department of Health, Albany.
Macario
A J
AJ
Mok
T
T
Beveridge
T J
TJ
D000907:Antibodies, Bacterial / Q000276:immunology; D000917:Antibody Formation; D000918:Antibody Specificity; D019843:Archaeal Proteins; D005455:Fluorescent Antibody Technique; D006026:Glycoside Hydrolases / Q000276:immunology* / Q000302:isolation & purification*; D007124:Immunoenzyme Techniques; D007150:Immunohistochemistry / Q000379:methods; D017020:Methanosarcina / Q000201:enzymology* / Q000648:ultrastructure; D016253:Microscopy, Immunoelectron / Q000379:methods; D008970:Molecular Weight
D000907:Antibodies, Bacterial; D019843:Archaeal Proteins; C067993:DAG protein, Methanosarcina mazei; D006026:Glycoside Hydrolases