Hemocyanin subunit organization of the gastropod Rapana thomasiana.
RtH1 and RtH2, the two hemocyanin isoforms of the prosobranch gastropod Rapana thomasiana, have been purified by anion-exchange chromatography and studied by SDS-PAGE and immunoelectrophoresis. Both subunit types are built up of eight functional units (FUs). Under reducing conditions subunit RtH2 splits into two fragments, RtH2-a-f and RtH2-gh, suggesting the presence of a disulfide bridge between FU2-f and FU2-g. By proteolytic cleavage of the subunits into three-, two-, and single-FU fragments, purification of fragments by HPLC, N-terminal sequencing of the peptides, and crossed-line immunoelectrophoresis, FUs-a-h of RtH2 and FU-a, FU-d, FU-e, and FU-f of RtH1 were identified and correlated to the eight-FUs pattern of immunoelectrophoresis. FU-a, FU-e, and FU-f of RtH1 and RtH2 are very closely related immunologically. RtH1 and RtH2 both correspond immunologically to KLH2, one of the two hemocyanin isoforms of the prosobranch gastropod Megathura crenulata.
Gebauer
W
W
Institute of Zoology, University of Mainz, Mainz, Germany. wolfgang@uzomai.biologie.uni-mainz.de
Stoeva
S
S
Voelter
W
W
Dainese
E
E
Salvato
B
B
Beltramini
M
M
Markl
J
J
D000595:Amino Acid Sequence; D000818:Animals; D006433:Hemocyanins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D007120:Immunochemistry; D007123:Immunoelectrophoresis, Two-Dimensional; D008969:Molecular Sequence Data; D008974:Mollusca / Q000737:chemistry* / Q000235:genetics; D010196:Pancreatic Elastase; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D020033:Protein Isoforms / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D020836:Protein Structure, Quaternary
D010446:Peptide Fragments; D020033:Protein Isoforms; D006433:Hemocyanins; D010196:Pancreatic Elastase