Carbohydrate gluing, an architectural mechanism in the supramolecular structure of an annelid giant hemoglobin.
We report a carbohydrate-dependent supramolecular architecture in the extracellular giant hemoglobin (Hb) from the marine worm Perinereis aibuhitensis; we call this architectural mechanism carbohydrate gluing. This study is an extension of our accidental discovery of deterioration in the form of the Hb caused by a high concentration of glucose. The giant Hbs of annelids are natural supramolecules consisting of about 200 polypeptide chains that associate to form a double-layered hexagonal structure. This Hb has 0.5% (wt) carbohydrates, including mannose, xylose, fucose, galactose, glucose, N-acetylglucosamine (GlcNAc), and N-acetylgalactosamine (GalNAc). Using carbohydrate-staining assays, in conjunction with two-dimensional polyacrylamide gel electrophoresis, we found that two types of linker chains (L1 and L2; the nomenclature of the Hb subunits followed that for another marine worm, Tylorrhynchus heterochaetus) contained carbohydrates with both GlcNAc and GalNAc. Furthermore, two types of globins (a and A) have only GlcNAc-containing carbohydrates, whereas the other types of globins (b and B) had no carbohydrates. Monosaccharides including mannose, fucose, glucose, galactose, GlcNAc, and GalNAc reversibly dissociated the intact form of the Hb, but the removal of carbohydrate with N-glycanase resulted in irreversible dissociation. These results show that carbohydrate acts noncovalently to glue together the components to yield the complete quaternary supramolecular structure of the giant Hb. We suggest that this carbohydrate gluing may be mediated through lectin-like carbohydrate-binding by the associated structural chains ("linkers").
Ebina
S
S
Nagayama Protein Array Project, Research Development Corporation of Japan, Tsukuba.
Matsubara
K
K
Nagayama
K
K
Yamaki
M
M
Gotoh
T
T
D000595:Amino Acid Sequence; D000818:Animals; D002241:Carbohydrates / Q000737:chemistry*; D006454:Hemoglobins / Q000737:chemistry* / Q000235:genetics / Q000648:ultrastructure; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D015394:Molecular Structure; D011077:Polychaeta / Q000737:chemistry* / Q000235:genetics; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid
D002241:Carbohydrates; D006454:Hemoglobins