Regulation of selectin binding activity by cyclization of sialic acid moiety of carbohydrate ligands on human leukocytes.
We provide here evidence that supports the occurrence of a biologically dormant form of selectin ligand carbohydrate, the sialyl 6-sulfo Lewis X containing modified sialic acid, in human leukocytes. The modification of sialic acid involves first de-N-acetylation of sialic acid moiety through ubiquitous de-N-acetylation/re-N-acetylation cycle, followed by the dehydrative cyclization of de-N-acetyl sialic acid to form "cyclic sialic acid." The enzyme involved in the dehydration of de-N-acetyl sialic acid is a calcium-dependent enzyme having neutral-alkaline pH optimum. De-N-acetyl sialyl 6-sulfo Lewis X retained selectin binding activity as well as parental sialyl 6-sulfo Lewis X, but cyclic sialyl 6-sulfo Lewis X was devoid of selectin binding activity. Sialyl 6-sulfo Lewis X carrying the cyclic sialic acid is specifically recognized by the newly generated mAb, G159. The determinant was distributed widely among normal human leukocytes, especially on monocytes and subsets of lymphocytes including NK cells, helper memory T cells, Tcr-gammadelta T cells, and a part of B cells. The determinant was detected also on several cultured lymphocytic leukemia cell lines and O-tetradecanoylphorbol 13-acetate-activated lymphoid cells. Cyclic sialyl 6-sulfo Lewis X is efficiently formed by the action of the partly membrane-bound calcium-dependent enzyme, tentatively called "sialic acid cyclase," and a possible physiological significance of this reaction could be a rapid inactivation of selectin binding activity at the cell surface. Conversely, the accumulated intracellular cyclic sialyl 6-sulfo Lewis X determinant may function as a dormant pool of selectin ligands, which, on appropriate stimulation, is hydrolyzed and becomes active in selectin-dependent cell adhesion.
Mitsuoka
C
C
Program of Experimental Pathology, Aichi Cancer Center, Nagoya 464-8681, Japan.
Ohmori
K
K
Kimura
N
N
Kanamori
A
A
Komba
S
S
Ishida
H
H
Kiso
M
M
Kannagi
R
R
D015703:Antigens, CD / Q000097:blood; D001665:Binding Sites; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002448:Cell Adhesion; D005434:Flow Cytometry; D006801:Humans; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D007700:Kinetics; D019041:L-Selectin / Q000378:metabolism*; D007962:Leukocytes / Q000502:physiology*; D016256:Lewis X Antigen / Q000031:analogs & derivatives; D008024:Ligands; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000097:blood* / Q000737:chemistry; D011994:Recombinant Proteins / Q000378:metabolism; D000080506:Sialyl Lewis X Antigen / Q000031:analogs & derivatives; D013329:Structure-Activity Relationship