Interactions of signaling proteins, growth factors and other proteins with heparan sulfate: mechanisms and mysteries

Abstract

Heparan sulfate (HS) is a component of cell surface and matrix-associated proteoglycans (HSPGs) that, collectively, play crucial roles in many physiologic processes including cell differentiation, organ morphogenesis and cancer. A key function of HS is to bind and interact with signaling proteins, growth factors, plasma proteins, immune-modulators and other factors. In doing so, the HS chains and HSPGs are able to regulate protein distribution, bio-availability and action on target cells and can also serve as cell surface co-receptors, facilitating ligand-receptor interactions. These proteins contain an HS/heparin-binding domain (HBD) that mediates their association and contacts with HS. HBDs are highly diverse in sequence and predicted structure, contain clusters of basic amino acids (Lys and Arg) and possess an overall net positive charge, most often within a consensus Cardin-Weintraub (CW) motif. Interestingly, other domains and residues are now known to influence protein-HS interactions, as well as interactions with other glycosaminoglycans, such as chondroitin sulfate. In this review, we provide a description and analysis of HBDs in proteins including amphiregulin, fibroblast growth factor family members, heparanase, sclerostin and hedgehog protein family members. We discuss HBD structural and functional features and important roles carried out by other protein domains, and also provide novel conformational insights into the diversity of CW motifs present in Sonic, Indian and Desert hedgehogs. Finally, we review progress in understanding the pathogenesis of a rare pediatric skeletal disorder, Hereditary Multiple Exostoses (HME), characterized by HS deficiency and cartilage tumor formation. Advances in understanding protein-HS interactions will have broad implications for basic biology and translational medicine as well as for the development of HS-based therapeutics.

Keywords: Extracellular matrix; heparan sulfate proteoglycans; heparan sulfate/heparin-binding domains; signaling and growth factor proteins.

Figure 1

Differential binding of AREG peptide to different GAGs. The indicated type of GAG was immobilized on 96 well plates and the binding of Flag- tagged AREG (DYKDDDDKGG RKKKGGKNGKNRRNRKKKN; AREG sequence underlined) peptide was determined using an anti-Flag Ab and secondary antibody-HPR conjugate. Results included are from a representative experiment and were highly reproducible.

Figure 2

Structure of CW motifs in Shh, Ihh and Dhh. Secondary structure predications were carried out using the I-TASSER server () and resulting structures were visualized using PyMol. The peptides are oriented with the N-terminus on the left; side chains of basic residues within the CW motifs are in blue while the side groups of proline are in red. Note that CWs of Shh and Ihh have a largely random coil configuration with central kinks due to the proline residue(s) while the CW of Dhh exhibits a more helical configuration. The sequences examined are presented below each structure; the CW motif is in bold type and flanking residues are in plain typeset.