Immunocytochemical localization of the multicatalytic proteinase (proteasome) in crustacean striated muscles.
Multicatalytic proteinase (MCP) is thought to play a central role in the processing and turnover of intracellular proteins in eukaryotic cells. Immunocytochemistry was used to determine the intracellular distribution of the MCP in the claw muscles of the land crab, Gecarcinus lateralis, and the claw and abdominal muscles of the American lobster, Homarus americanus. Cryosections were stained with an affinity-purified polyclonal antibody to lobster MCP that cross-reacted with the land crab enzyme. Two types of staining were observed: a diffuse cytoplasmic staining, and a dense aggregate staining primarily associated with invaginations of the cell membrane. The cytoplasmic staining appeared reticulated in favorable transverse sections due to a preferential localization of MCP to the intermyofibrillar space. The aggregate staining was associated with neither nuclei nor mitochondria, since stains specific for these organelles (Hoechst stain and nicotinamide adenine dinucleotide diaphorase histochemistry, respectively) did not colocalize with the aggregates. Trypsinlike peptidase activities of isolated microsomal and postmicrosomal fractions indicated that less than 1% of the total MCP was associated with the microsomal fraction. Immunoprecipitation of the same fractions confirmed the presence of MCP in the microsomes as well as in the cytosol. These results suggest that the MCP is primarily associated with cytoplasmic components; the aggregate staining may result from the association of the MCP with cellular membrane systems.
Beyette
J R
JR
Department of Biology, Colorado State University, Fort Collins 80523.
Mykles
D L
DL
D000818:Animals; D003386:Brachyura / Q000201:enzymology*; D002462:Cell Membrane / Q000737:chemistry; D003546:Cysteine Endopeptidases / Q000032:analysis*; D015151:Immunoblotting; D007150:Immunohistochemistry; D008861:Microsomes / Q000737:chemistry; D009097:Multienzyme Complexes / Q000032:analysis*; D009132:Muscles / Q000201:enzymology*; D008121:Nephropidae / Q000201:enzymology*; D046988:Proteasome Endopeptidase Complex; D011817:Rabbits; D012508:Sarcolemma / Q000737:chemistry
D009097:Multienzyme Complexes; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex