Cryo-EM structures of human A2ML1 elucidate the protease-inhibitory mechanism of the A2M family

• Published in: Nature communications Vol. 13; no. 1; p. 3033
• Main Authors: Nielsen, Nadia Sukusu, Zarantonello, Alessandra, Harwood, Seandean Lykke, Jensen, Kathrine Tejlgård, Kjøge, Katarzyna, Thøgersen, Ida B., Schauser, Leif, Karlsen, Jesper Lykkegaard, Andersen, Gregers R., Enghild, Jan J.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 31.05.2022; Nature Publishing Group; Nature Portfolio
• Subjects: 101/28; 101/58; 631/250/2501; 631/45/468; 631/535/1258/1259; 82/80; 82/81; 82/83; alpha-Macroglobulins - chemistry; alpha-Macroglobulins - metabolism; Baits; Complement component C3; Complement component C4; Cryoelectron Microscopy; Endopeptidases; Humanities and Social Sciences; Humans; Hydrophobicity; multidisciplinary; Peptide Hydrolases; Protease; Protease inhibitors; Protease Inhibitors - pharmacology; Proteinase inhibitors; Proteins; Proteolysis; Science; Science (multidisciplinary); Substrates
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-022-30758-x

A2ML1 is a monomeric protease inhibitor belonging to the A2M superfamily of protease inhibitors and complement factors. Here, we investigate the protease-inhibitory mechanism of human A2ML1 and determine the structures of its native and protease-cleaved conformations. The functional inhibitory unit of A2ML1 is a monomer that depends on covalent binding of the protease (mediated by A2ML1’s thioester) to achieve inhibition. In contrast to the A2M tetramer which traps proteases in two internal chambers formed by four subunits, in protease-cleaved monomeric A2ML1 disordered regions surround the trapped protease and may prevent substrate access. In native A2ML1, the bait region is threaded through a hydrophobic channel, suggesting that disruption of this arrangement by bait region cleavage triggers the extensive conformational changes that result in protease inhibition. Structural comparisons with complement C3/C4 suggest that the A2M superfamily of proteins share this mechanism for the triggering of conformational change occurring upon proteolytic activation. A2ML1 is a human protease inhibitor belonging to the A2M protein family. In this study, the authors determine structures of A2ML1 before and after protease inhibition and investigate its mechanism of action.