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.
Kozhemyako
Valeri B
VB
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
Rebrikov
Denis V
DV
Lukyanov
Sergey A
SA
Bogdanova
Ekaterina A
EA
Marin
Antoine
A
Mazur
Alexey K
AK
Kovalchuk
Svetlana N
SN
Agafonova
Elena V
EV
Sova
Victoria V
VV
Elyakova
Ludmila A
LA
Rasskazov
Valeri A
VA
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