TY - JOUR
T1 - Hemocyanin subunit organization of the gastropod Rapana thomasiana
AU - Gebauer, Wolfgang
AU - Stoeva, Stanka
AU - Voelter, Wolfgang
AU - Dainese, Enrico
AU - Salvato, Benedetto
AU - Beltramini, Mariano
AU - Markl, Jürgen
PY - 1999/12/1
Y1 - 1999/12/1
N2 - 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-α-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-α-h of RtH2 and FU-α, FU-d, FU-e, and FU-f of RtH1 were identified and correlated to the eight-FUs pattern of immunoelectrophoresis. FU-α, 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.
AB - 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-α-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-α-h of RtH2 and FU-α, FU-d, FU-e, and FU-f of RtH1 were identified and correlated to the eight-FUs pattern of immunoelectrophoresis. FU-α, 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.
KW - Disulfide bridge
KW - Hemocyanin
KW - Mollusca
KW - Rapana thomasiana
KW - Subunit structure
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U2 - 10.1006/abbi.1999.1478
DO - 10.1006/abbi.1999.1478
M3 - Article
C2 - 10562425
AN - SCOPUS:0033485297
VL - 372
SP - 128
EP - 134
JO - Archives of Biochemistry and Biophysics
JF - Archives of Biochemistry and Biophysics
SN - 0003-9861
IS - 1
ER -