Horseshoe crab acetyl group-recognizing lectins involved in innate immunity are structurally related to fibrinogen

TLDR

The horseshoe crab possesses a fibrinogen‑like coagulogen that may reflect an ancestral nonself‑recognizing protein. We cloned two lectins, TLs‑5A and TLs‑5B, from horseshoe crab hemolymph that agglutinate human red cells and bacteria, recognize acetyl groups, enhance big defensin activity, and possess a fibrinogen‑like C‑terminal domain forming propeller structures, indicating they are nonself‑recognizing lectins.

Abstract

We have characterized and cloned newly isolated lectins from hemolymph plasma of the horseshoe crab Tachypleus tridentatus , which we named tachylectins 5A and 5B (TLs-5). TLs-5 agglutinated all types of human erythrocytes and Gram-positive and Gram-negative bacteria. TLs-5 specifically recognize acetyl group-containing substances including noncarbohydrates; the acetyl group is required and is sufficient for recognition. TLs-5 enhanced the antimicrobial activity of a horseshoe crab-derived big defensin. cDNA sequences of TLs-5 indicated that they consist of a short N-terminal Cys-containing segment and a C-terminal fibrinogen-like domain with the highest sequence identity (51%) to that of mammalian ficolins. TLs-5, however, lack the collagenous domain found in a kind of “bouquet arrangement” of ficolins and collectins. Electron microscopy revealed that TLs-5 form two- to four-bladed propeller structures. The horseshoe crab is equipped with a unique functional homologue of vertebrate fibrinogen, coagulogen, as the target protein of the clotting cascade. Our observations clearly show that the horseshoe crab has fibrinogen-related molecules in hemolymph plasma and that they function as nonself-recognizing lectins. An ancestor of fibrinogen may have functioned as a nonself-recognizing protein.