PMID- 10393093
OWN - NLM
STAT- MEDLINE
DCOM- 19990908
LR  - 20181113
IS  - 0264-6021 (Print)
IS  - 0264-6021 (Linking)
VI  - 341 ( Pt 2)
DP  - 1999 Jul 15
TI  - Molecular analysis of sialoside binding to sialoadhesin by NMR and site-directed 
      mutagenesis.
PG  - 355-61
AB  - The molecular interactions between sialoadhesin and sialylated ligands have been 
      investigated by using proton NMR. Addition of ligands to the 12 kDa N-terminal
      immunoglobulin-like domain of sialoadhesin result in resonance shifts in the
      protein spectrum that have been used to determine the affinities of sialoadhesin 
      for several sialosides. The results indicate that alpha2, 3-sialyl-lactose and
      alpha2,6-sialyl-lactose bind respectively 2- and 1.5-fold more strongly than does
      alpha-methyl-N-acetylneuraminic acid (alpha-Me-NeuAc). The resonances
      corresponding to the methyl protons within the N-acetyl moiety of sialic acid
      undergo upfield shifting and broadening during titrations, reflecting an
      interaction of this group with Trp2 in sialoadhesin as observed in co-crystals of
      the terminal domain with bound ligand. This resonance shift was used to measure
      the affinities of mutant and wild-type forms of sialoadhesin in which the first
      three domains are fused to the Fc region of human IgG1. Substitution of Arg97 by 
      alanine completely abrogated measurable interaction with alpha-Me-NeuAc, whereas 
      a conservative substitution with lysine resulted in a 10-fold decrease in
      affinity. These results provide the first direct measurement of the affinity of
      sialoadhesin for sialosides and confirm the critical importance of the conserved 
      arginine in interactions between sialosides and members of the siglec family of
      sialic acid-binding, immunoglobulin-like lectins.
FAU - Crocker, P R
AU  - Crocker PR
AD  - Department of Biochemistry, Wellcome Trust Building, University of Dundee, Dundee
      DD1 5EH, Scotland, U.K.
FAU - Vinson, M
AU  - Vinson M
FAU - Kelm, S
AU  - Kelm S
FAU - Drickamer, K
AU  - Drickamer K
LA  - eng
GR  - Wellcome Trust/United Kingdom
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Biochem J
JT  - The Biochemical journal
JID - 2984726R
RN  - 0 (Ligands)
RN  - 0 (Membrane Glycoproteins)
RN  - 0 (Receptors, Immunologic)
RN  - 0 (SIGLEC1 protein, human)
RN  - 0 (Sialic Acid Binding Ig-like Lectin 1)
RN  - 0 (Sialic Acids)
RN  - 3J7TAL60G3 (N-acetylneuraminoyllactose)
RN  - 67974-39-4 (4-O-methyl-N-acetylneuraminic acid)
RN  - J2B2A4N98G (Lactose)
SB  - IM
MH  - Binding Sites
MH  - Humans
MH  - Lactose/*analogs & derivatives/chemistry/genetics/metabolism
MH  - Ligands
MH  - Magnetic Resonance Spectroscopy
MH  - Membrane Glycoproteins/chemistry/genetics/*metabolism
MH  - Mutagenesis, Site-Directed
MH  - Receptors, Immunologic/chemistry/genetics/*metabolism
MH  - Sialic Acid Binding Ig-like Lectin 1
MH  - Sialic Acids/chemistry/genetics/*metabolism
PMC - PMC1220367
EDAT- 1999/07/07 00:00
MHDA- 1999/07/07 00:01
CRDT- 1999/07/07 00:00
PHST- 1999/07/07 00:00 [pubmed]
PHST- 1999/07/07 00:01 [medline]
PHST- 1999/07/07 00:00 [entrez]
PST - ppublish
SO  - Biochem J. 1999 Jul 15;341 ( Pt 2):355-61.