Molecular cloning and characterization of an endo-1,3-beta-D-glucanase from the mollusk Spisula sachalinensis.
cDNA encoding the endo-1,3-beta-d-glucanase from Spisula sachalinensis (LIV) was amplified by PCR using oligonucleotides deduced from the N-terminal end peptide sequence. Predicted enzyme structure consists of 444 amino acids with a signal sequence. The mature enzyme has 316 amino acids and its deduced amino acid sequence coincides completely with the N-terminal end (38 amino acids) of the beta-1,3-glucanase (LIV) isolated from the mollusk. The enzyme sequence from Val 121 to Met 441 reveals closest homology with Pacifastacus leniusculus lipopolysaccharide- and beta-1,3-glucan-binding protein and with coelomic cytolytic factors from Lumbricus terrestris. The mollusk glucanase also shows 36% identity and 56% similarity with beta-1,3-glucanase of the sea urchin Strongylocentrotus purpuratus. It is generally considered that invertebrate glucanase-like proteins containing the bacterial glucanase motif have evolved from an ancient beta-1,3-glucanase gene, but most of them lost their glucanase activity in the course of evolution and retained only the glucan-binding activity. A more detailed evaluation of the protein folding elicited very interesting relationships between the active site of LIV and other enzymes, which hydrolyze native glucans.
Pacific Institute of Bioorganic Chemistry, Far East Branch of the Russian Academy of Sciences, pr.100 let Vladivostoku, 159, 690022, Vladivostok, Russia. kozhemyako@piboc.dvo.ru
Valeri B
VB
Kozhemyako
Denis V
DV
Rebrikov
Sergey A
SA
Lukyanov
Ekaterina A
EA
Bogdanova
Antoine
A
Marin
Alexey K
AK
Mazur
Svetlana N
SN
Kovalchuk
Elena V
EV
Agafonova
Victoria V
VV
Sova
Ludmila A
LA
Elyakova
Valeri A
VA
Rasskazov
D000595:Amino Acid Sequence; D000818:Animals; D003400:Astacoidea / Q000235:genetics; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004693:Glucan Endo-1,3-beta-D-Glucosidase / Q000235:genetics*; D008969:Molecular Sequence Data; D008974:Mollusca / Q000235:genetics*; D010802:Phylogeny; D017434:Protein Structure, Tertiary; D012617:Sea Urchins / Q000235:genetics; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; D004693:Glucan Endo-1,3-beta-D-Glucosidase