Neurotrophin binding to human alpha 2-macroglobulin under apparent equilibrium conditions

Biochemistry
B B Wolf, S L Gonias

Abstract

alpha 2-Macroglobulin (alpha 2M) is a broad-spectrum proteinase inhibitor and a carrier of certain growth factors. The purpose of this investigation was to characterize the interaction of alpha 2M with nerve growth factor-beta (NGF-beta), brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), neurotrophin-4 (NT-4), and ciliary neurotrophic factor (CNTF) under apparent equilibrium conditions. Binding in solution was assessed using the cross-linking agent bis(sulfosuccinimidyl) suberate (BS3). Noncovalent binding of NGF-beta, NT-3, NT-4, and BDNF to native alpha 2M and alpha 2M-methylamine (a conformationally modified form of alpha 2M that is recognized by the alpha 2M receptor) reached apparent equilibrium in less than 20 min at 37 degrees C. Apparent KD values for the binding of NT-4, NGF-beta, NT-3, and BDNF to alpha 2M-methylamine were 61, 110, 120, and 150 nM, respectively. Native alpha 2M bound all four neurotrophins with decreased affinity. Unlabeled NGF-beta competed with the radioiodinated neurotrophins for binding to immobilized alpha 2M-methylamine. The K1 for unlabeled NGF-beta was 120 nM, in good agreement with the apparent KD determined by the BS3 method. The number of NGF-beta binding sites per immobilize...Continue Reading

Citations

May 24, 2008·The Journal of Biological Chemistry·Elisabetta MantuanoSteven L Gonias
Aug 15, 2008·The Journal of Biological Chemistry·ZhiHua ShiH Uri Saragovi
Mar 14, 2000·Journal of Interferon & Cytokine Research : the Official Journal of the International Society for Interferon and Cytokine Research·T R GarberD J Webb
Mar 3, 1999·Proceedings of the National Academy of Sciences of the United States of America·G VenkataramanR Sasisekharan
May 15, 2007·Brain Research Bulletin·Viktorija TrajkovskaGitte M Knudsen
Feb 4, 2011·Journal of Neuroscience Research·Yang ShiSteven L Gonias
Oct 23, 2012·Journal of Cellular Physiology·Ahmed A RehmanFahim H Khan
Nov 5, 1999·Analytical Biochemistry·C E MacPheeW H Sawyer
Aug 21, 2019·Oxidative Medicine and Cellular Longevity·Jordan H CaterAmy R Wyatt
Dec 17, 2020·Frontiers in Immunology·Jessica Le BlancMarie Lordkipanidzé

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