The interaction of biological molecules with heparin and related glycosaminoglycans. 1. Identification of a specific heparin binding site for histamine

Abstract

The interaction of histamine with beef lung heparin has been characterized by 'H and I3C NMR spectroscopy. IH chemical shifts provide evidence that histamine forms a complex with heparin both at low pD where 2-Osulfo-a-~-idopyranosyluronic acid (IdoA-2s) residues are in the carboxylic acid form and at higher pD where the carboxyl groups are deprotonated. At low pD, the chemical shift data are consistent with weak delocalized binding of the diprotonated form of histamine by the highly negatively charged polymer. However, as the pD is increased and the carboxylic acid groups are titrated, a binding region with a high affinity for diprotonated histamine is created. It is proposed that, in this binding site, the ammonium group of histamine is hydrogen-bonded to the carboxylate group of an IdoA-2S residue while the imidazolium ring is located in a region surrounded on three sides by the sulfamido group of a 2-deoxy-2-sulfamido-6.Osulfo-a-Dglucopyr (GlcNS03-6S) residue and the 2-Osulfate and carboxylate groups of a second IdoA-2S residue. Evidence for site specific binding includes displacement of chemical shift titration curves to lower pD and increased shielding of specific heparin protons from the imidazole ring current. Similar 'H NMR measurements on the heparin-imidazole and heparin-N-acetylhistamine systems indicate that the highly negatively charged binding site created by deprotonation of the carboxylic acid groups has a high affinity in general for positively charged imidazolium rings. Nuclear Overhauser enhancements obtained from two-dimensional NOESY spectra of heparin at pD 2.4 and 7.4 indicate that, when the carboxylic acid groups of heparin are titrated, its conformation changes by a small rotation around the (IdoA-2S)-( 1-+4)-(GlcNS03-6S) glycosidic bond. This study provides evidence for the first time of the site specific binding of a biological molecule by the repeating (IdoA-2S)-( 1+4)-(GlcNS03-6s) disaccharide unit of heparin.

CD of proline‐rich polypeptides: Application to the study of the repetitive domain of maize glutelin‐2

“…Recently, an accurate nmr study dealing with the interaction of histamine to heparin has established that the binding of histamine to heparin is site specific, i-e., the ammonium group binds to the carboxylate group, whereas the imidazolium ring is located in a pocket surrounded by three anionic groups, two 0-sulfate groups, and the carboxylate of the following ring. 42 The finding of a binding site Structure of the repeating disaccharide unit with high affinity for the imidazolium ring is of particular interest for the interaction of heparine with histidine-containing peptides, 43 as in the present article.…”

Binding of the growth factor glycyl‐l‐histidyl‐l‐lysine by heparin

“…shown below with the iduronic acid residue (the I ring) in the ~C4 conformation and the glucosamine (the A ring) in the 4C t conformation. Because of its high negative charge density, many biological molecules, including histamine and many peptides and proteins [15,17], bind to heparin. The binding mechanism may be either delocalized (territorial) or site specific electrostatic interactions between anionic sites on heparin and cationic sites on the biological molecules [15].…”

“…In this paper, we report that GHK binds to the glycosaminoglycan heparin; the binding interaction has been characterized by [IH]NMR spectroscopy and is shown to involve the Nterminal glycyl ammonium, the histidyl imidazolium and the lysine ammonium groups, with the imidazolium group interacting site specifically with the imidazolium-binding site on heparin [17].…”

Estimating the Time Course of Pore Expansion During the Spike Phase of Exocytotic Release in Mast Cells of the Beige Mouse

“…We examined the possibility that it could represent the first stages of 'resealing' of the cell (Nielsen et al, 1981;Steyer et al, 1997), but found that pore contraction caused the efflux to decrease at a faster, not a slower rate. There is a difference of about 1.2 pH units between the interior of the vesicle (pH 6.0) and extracellular solution (Johnson et al, 1980) and since binding to heparin, at least for histamine, depends upon pH (Rabenstein et al, 1992) it is expected that D will vary during the release process as the pH gradient equilibrates. In addition, there is evidence that D would increase (not decrease) by about a factor of 2 as the gel expands and the network becomes less tortuous (Krizaj et al, 1996;Rusakov and Kullmann, 1998).…”