Probing sialic acid binding Ig-like lectins (siglecs) with sulfated oligosaccharides.
Soluble siglecs-1, -4, -5, -6, -7, -8, -9, and -10 were probed with polyacrylamide glycoconjugates in which: 1) the Neu5Ac residue was substituted by a sulfate group (Su); 2) glycoconjugates contained both Su and Neu5Ac; 3) sialoglycoconjugates contained a tyrosine-O-sulfate residue. It was shown that sulfate derivatives of LacNAc did not bind siglecs-1, -4, -5, -6, -7, -8, -9, and -10; binding of 6'-O-Su-LacNAc to siglec-8 was stronger than binding of 3'SiaLacNAc. The relative affinity of 3'-O-Su-TF binding to siglecs-1, -4, and -8 was similar to that of 3'SiaTF. 3'-O-Su-Le(c) displayed two-fold weaker binding to siglec-1 and siglec-4 than 3'SiaLe(c). The interaction of soluble siglecs with sulfated oligosaccharides containing sialic acid was also studied. It was shown that siglecs-1, -4, -5, -6, -7, -9, and -10 did not interact with these compounds; binding of 6-O-Su-3'SiaLacNAc and 6-O-Su-3'SiaTF to siglec-8 was weaker than that of the corresponding sulfate-free sialoside probes. Siglec-8 displayed affinity to 6'-O-Su-LacNAc and 6'-O-Su-SiaLe(x), and defucosylation of the latter compound led to an increase in the binding. Sialoside probes containing tyrosine-O-sulfate residue did not display increased affinity to siglecs-1 and -5 compared with glycoconjugates containing only sialoside. Cell-bound siglecs-1, -5, -7, and -9 did not interact with 6-O-Su-3'SiaLacNAc, whereas the sulfate-free probe 3'SiaLacNAc demonstrated binding. In contrast, the presence of sulfate in 6-O-Su-6'SiaLacNAc did not affect binding of the sialoside probe to siglecs. 6'-O-Su-SiaLe(x) displayed affinity to cell-bound siglecs-1 and -5; its isomer 6-O-Su-SiaLe(x) bound more strongly to siglecs-1, -5, and -9 than SiaLe(x).
Rapoport
E M
EM
Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russia.
Pazynina
G V
GV
Sablina
M A
MA
Crocker
P R
PR
Bovin
N V
NV
D000818:Animals; D015703:Antigens, CD / Q000235:genetics / Q000378:metabolism; D015214:Antigens, Differentiation, Myelomonocytic / Q000235:genetics / Q000378:metabolism; D016466:CHO Cells; D002240:Carbohydrate Sequence; D006224:Cricetinae; D003412:Cricetulus; D006001:Glycoconjugates / Q000737:chemistry / Q000494:pharmacology; D006801:Humans; D037102:Lectins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000037:antagonists & inhibitors; D051379:Mice; D009844:Oligosaccharides / Q000737:chemistry*; D011971:Receptors, Immunologic / Q000037:antagonists & inhibitors; D063267:Sialic Acid Binding Ig-like Lectin 1; D063265:Sialic Acid Binding Immunoglobulin-like Lectins; D012794:Sialic Acids / Q000378:metabolism; D013379:Substrate Specificity; D013431:Sulfates / Q000737:chemistry*; D014443:Tyrosine / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism
D015703:Antigens, CD; D015214:Antigens, Differentiation, Myelomonocytic; D006001:Glycoconjugates; D037102:Lectins; D008562:Membrane Glycoproteins; D009844:Oligosaccharides; D011971:Receptors, Immunologic; C571166:SIGLEC1 protein, human; C114531:SIGLEC5 protein, human; C402206:SIGLEC7 protein, human; C412227:SIGLEC9 protein, human; D063267:Sialic Acid Binding Ig-like Lectin 1; D063265:Sialic Acid Binding Immunoglobulin-like Lectins; D012794:Sialic Acids; C571167:Siglec1 protein, mouse; D013431:Sulfates; C005662:tyrosine O-sulfate; D014443:Tyrosine