Antibodies to mammalian and plant V-ATPases cross react with the V-ATPase of insect cation-transporting plasma membranes.
In immunobiochemical blots, polyclonal antibodies against subunits of plant and mammalian vacuolar-type ATPases (V-ATPases) cross-react strongly with corresponding subunits of larval Manduca sexta midgut plasma membrane V-ATPase. Thus, rabbit antiserum against Kalanchoe daigremontiana tonoplast V-ATPase holoenzyme cross-reacts with the 67, 56, 40, 28 and 20 kDa subunits of midgut V-ATPase separated by SDS-PAGE. Antisera against bovine chromaffin granule 72 and 39 kDa V-ATPase subunits cross-react with the corresponding 67 and 43 kDa subunits of midgut V-ATPase. Antisera against the 57 kDa subunit of both beet root and oat root V-ATPase cross-react strongly with the midgut 56 kDa V-ATPase subunit. In immunocytochemical light micrographs, antiserum against the beet root 57 kDa V-ATPase subunit labels the goblet cell apical membrane of both posterior and anterior midgut in freeze-substituted and fixed sections. The plant antiserum also labels the apical brush-border plasma membrane of Malpighian tubules. The ability of antibodies against plant V-ATPase to label these insect membranes suggests a high sequence homology between V-ATPases from plants and insects. Both of the antibody-labelled insect membranes transport K+ and both membranes possess F1-like particles, portasomes, on their cytoplasmic surfaces. This immunolabelling by xenic V-ATPase antisera of two insect cation-transporting membranes suggests that the portasomes on these membranes may be V-ATPase particles, similar to those reported on V-ATPase-containing vacuolar membranes from various sources.
Russell
V E
VE
Department of Entomology, Purdue University, West Lafayette, IN 47907.
Klein
U
U
Reuveni
M
M
Spaeth
D D
DD
Wolfersberger
M G
MG
Harvey
W R
WR
D000251:Adenosine Triphosphatases / Q000276:immunology*; D000818:Animals; D000906:Antibodies; D001693:Biological Transport, Active; D002412:Cations; D002417:Cattle; D002462:Cell Membrane / Q000201:enzymology; D002838:Chromaffin System / Q000201:enzymology; D003429:Cross Reactions; D004064:Digestive System / Q000201:enzymology; D007120:Immunochemistry; D008317:Malpighian Tubules / Q000201:enzymology; D009036:Moths / Q000201:enzymology*; D010944:Plants / Q000201:enzymology*; D013045:Species Specificity; D014617:Vacuoles / Q000201:enzymology
D000906:Antibodies; D002412:Cations; D000251:Adenosine Triphosphatases