Nix Matthew A, Kaelin Christopher B, Palomino Rafael, Miller Jillian L, Barsh Gregory S, Millhauser Glenn L
Department of Chemistry and Biochemistry, University of California, Santa Cruz, Santa Cruz, California.
HudsonAlpha Institute for Biotechnology, Huntsville, Alabama; Department of Genetics, Stanford University, Stanford, California.
Biophys J. 2015 Nov 3;109(9):1946-58. doi: 10.1016/j.bpj.2015.09.005.
The β-defensins are a class of small cationic proteins that serve as components of numerous systems in vertebrate biology, including the immune and melanocortin systems. Human β-defensin 3 (HBD3), which is produced in the skin, has been found to bind to melanocortin receptors 1 and 4 through complementary electrostatics, a unique mechanism of ligand-receptor interaction. This finding indicates that electrostatics alone, and not specific amino acid contact points, could be sufficient for function in this ligand-receptor system, and further suggests that other small peptide ligands could interact with these receptors in a similar fashion. Here, we conducted molecular-similarity analyses and functional studies of additional members of the human β-defensin family, examining their potential as ligands of melanocortin-1 receptor, through selection based on their electrostatic similarity to HBD3. Using Poisson-Boltzmann electrostatic calculations and molecular-similarity analysis, we identified members of the human β-defensin family that are both similar and dissimilar to HBD3 in terms of electrostatic potential. Synthesis and functional testing of a subset of these β-defensins showed that peptides with an HBD3-like electrostatic character bound to melanocortin receptors with high affinity, whereas those that were anticorrelated to HBD3 showed no binding affinity. These findings expand on the central role of electrostatics in the control of this ligand-receptor system and further demonstrate the utility of employing molecular-similarity analysis. Additionally, we identified several new potential ligands of melanocortin-1 receptor, which may have implications for our understanding of the role defensins play in melanocortin physiology.
β-防御素是一类小的阳离子蛋白,在脊椎动物生物学的众多系统中发挥作用,包括免疫和黑皮质素系统。已发现皮肤中产生的人β-防御素3(HBD3)通过互补静电作用与黑皮质素受体1和4结合,这是一种独特的配体-受体相互作用机制。这一发现表明,仅静电作用而非特定的氨基酸接触点,就足以在该配体-受体系统中发挥功能,进一步表明其他小肽配体可能以类似方式与这些受体相互作用。在此,我们对人β-防御素家族的其他成员进行了分子相似性分析和功能研究,通过基于它们与HBD3的静电相似性进行筛选,来检验它们作为黑皮质素-1受体配体的潜力。使用泊松-玻尔兹曼静电计算和分子相似性分析,我们鉴定出在静电势方面与HBD3既有相似之处又有不同之处的人β-防御素家族成员。对这些β-防御素的一个子集进行合成和功能测试表明,具有HBD3样静电特征的肽与黑皮质素受体具有高亲和力结合,而那些与HBD3呈反相关的肽则没有结合亲和力。这些发现扩展了静电作用在控制该配体-受体系统中的核心作用,并进一步证明了采用分子相似性分析的实用性。此外,我们鉴定出了几种新的黑皮质素-1受体潜在配体,这可能对我们理解防御素在黑皮质素生理学中的作用具有启示意义。