Heavy metal binding to heparin disaccharides. I. Iduronic acid is the main binding site
Version of Record online: 1 FEB 2004
Copyright © 1992 John Wiley & Sons, Inc.
Volume 32, Issue 6, pages 585–596, June 1992
How to Cite
Whitfield, D. M., Choay, J. and Sarkar, B. (1992), Heavy metal binding to heparin disaccharides. I. Iduronic acid is the main binding site. Biopolymers, 32: 585–596. doi: 10.1002/bip.360320603
- Issue online: 1 FEB 2004
- Version of Record online: 1 FEB 2004
- Manuscript Accepted: 3 OCT 1991
- Manuscript Received: 4 JUN 1991
As model compounds for Ni(II)-binding heparin-like compounds isolated from human kidneys (Templeton, D. M. & Sarkar, B. (1985) Biochem. J.230 35–42.), we investigated two disaccharides—4-O-(2-O-sulfo-α-L-idopyranosyluronic acid)-2,5-anhydro-D-mannitol, disodium salt (1a), and 4-O-(2-O-sulfo-α-L-idopyranosyluronic acid)-6-O-sulfo-2,5-an-hydro-D-mannitol, trisodium salt (1b)—that were isolated from heparin after nitrous acid hydrolysis and reduction. The monosulfate (1a) was active whereas the disulfate (1b) was inactive in a high-performance liquid chromatography (HPLC) binding assay with the tracer ions 63Ni(II) 54Mn(II), 65Zn(II), and 109Cd(II). This result is in accord with the isolation of two 67Cu(II) and 63Ni(II) binding fractions from a complete pool of nitrous-acid-derived heparin disaccharides using sulfate gradients and a MonoQ anion exchange column on an FPLC system. One was identified as compound (1a) and the other as a tetrasulfated trisaccharide by high resolution FAB-MS, NMR and HPLC-PAD. Similarly, two synthetic disaccharides—methyl, 2-O-sulfo-4-O-(α-L-idopyranosyluronic acid)-2-deoxy-2-sulfamido-α-D-glucosamine, trisodium salt [IdopA2S(α1,4)GlcNSαMe,2a], and 2-O-sulfo-4-O- (α-L-idopyranosyluronic acid)-2-deoxy-2-sulfamide-6-O-sulfo-α-D-glucosamine, tetrasodium salt [IdopA2S(α1,4) GlcNS6SαMe,2b]—were shown to bind tracer amounts of 63Ni and 67Cu using chromatographic assays. Subsequently, 1H NMR titrations of 1a, 1b, 2a, and 2b with Zn(OAc)2 were analyzed to yield 1:1 Zn(II)-binding constants of 472 ± 59, 698 ± 120, 8,758 ± 2,237 and 20,100 ± 5,598M−1, respectively. The values for 2a and 2b suggest chelation. It is suggested that the idopyranosiduronic acid residue is the major metal binding site. NMR evidence for this hypothesis comes from marked 1H and 13C chemical shift changes to the iduronic acid resonances after addition of diamagnetic Zn(II) ions.