Hemocyanin subunit organization of the gastropod Rapana thomasiana

Citation
W. Gebauer et al., Hemocyanin subunit organization of the gastropod Rapana thomasiana, ARCH BIOCH, 372(1), 1999, pp. 128-134
Citations number
25
Categorie Soggetti
Biochemistry & Biophysics
Journal title
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS
ISSN journal
00039861 → ACNP
Volume
372
Issue
1
Year of publication
1999
Pages
128 - 134
Database
ISI
SICI code
0003-9861(199912)372:1<128:HSOOTG>2.0.ZU;2-N
Abstract
RtH1 and RtH2, the two hemocyanin isoforms of the prosobranch gastropod Rap ana thomasiana, have been purified by anion-exchange chromatography and stu died by SDS-PAGE and immunoelectrophoresis. Both subunit types are built up of eight functional units (FUs). Under reducing conditions subunit RtH2 sp lits 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 s ubunits into three-, two-, and single-FU fragments, purification of fragmen ts by HPLC, N-terminal sequencing of the peptides, and crossed-line immunoe lectrophoresis, 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 immunoelectrophoresi s. FU-a, FU-e, and FU-f of RtH1 and RtH2 are very closely related immunolog ically. RtH1 and RtH2 both correspond immunologically to KLH2, one of the t wo hemocyanin isoforms of the prosobranch gastropod Megathura crenulata. (C ) 1999 Academic Press.