Pulcherosine, a novel tyrosine-derived, trivalent crosslinking amino acid from the fertilization envelope of sea urchin embryo

Abstract

The normally hardened and aminotriazole-induced soft fertilization envelopes (FEs) of the sea urchin Hemicentrotus pulcherrimus and two other species were isolated and investigated for component proteins and cross-linking amino acids. From the acid hydrolysate of the hard FE of H. pulcherrimus, we isolated by reversed-phase high-performance liquid chromatography a novel fluorescent compound as well as dityrosine and trityrosine, the major tyrosine-derived cross-linking amino acids. These three compounds were also isolated from the reaction products of the tyrosine/horseradish peroxidase/H2O2 system. The structure of the novel compound, designated "pulcherosine", was determined to be 5-[4"-(2-carboxy-2-aminoethyl)phenoxy]-3,3'-dityrosine. With respect to the position of diphenyl ether bond between the tyrosine and dityrosine moieties, it is an isomer of isotrityrosine found in Ascaris cuticle collagen [Fujimoto et al. (1981) Biochem. Biophys. Res. Commun. 99, 637-643]. Isotrityrosine was not found in either of the above systems as a major component. The contents of tyrosine, dityrosine, trityrosine, and pulcherosine in the hard FE of H. pulcherrimus were estimated as 255, 5.5, 2.1, and 1.3 residues, respectively, per 10,000 total amino acid residues, while in the soft FE, those of tyrosine and dityrosine were 305 and 0.25 residues, respectively, and trityrosine and pulcherosine were only traces. The molar ratio of dityrosine, trityrosine, and pulcherosine in the hard FE was 100:38:24, while that for tyrosine/horseradish peroxidase/H2O2 reaction products was 100:3:8, respectively.

Human Phagocytes Employ the Myeloperoxidase-Hydrogen Peroxide System to Synthesize Dityrosine, Trityrosine, Pulcherosine, and Isodityrosine by a Tyrosyl Radical-dependent Pathway

“…The extinction coefficients, ultraviolet absorption spectra, and fluorescence excitation/emission spectra of the compounds were then determined at acid and alkaline pH. These data, which are in good agreement with previously reported results for tyrosine oxidation products isolated from sea urchins, insects, and yeast (24,25,32,36), are summarized in Table I.…”

“…This region of compound I ( Fig. 2A) was very similar to that previously reported for 3,3Ј-dityrosine isolated from yeast spore coats (32) To confirm the assignments of the protons in compound II, we used total correlation spectroscopy to examine the connectivities of the protons (Fig. 3).…”

“…Tyrosine oxidation products, including dityrosine (30,31), pulcherosine (32), isodityrosine (33), and trityrosine (30,32), are a common post-translational modification found in bacteria (34), yeast (35,36), plants (33), and metazoans (30 -33, 37). In * This work was supported in part by a grant-in-aid from the American Heart Association and by National Institutes of Health Grants RO1 HD29920 and RO1 AG12293.…”

Identification of an Intermediate State in the Helix-Coil Degradation of Collagen by Ultraviolet Light

“…Dityrosine was synthesized by peroxide oxidation of tyrosine according to the method described by Nomura et al (19).…”

Some Properties of the Hardening Process in Chorions Isolated from Unfertilized Eggs of Medaka, Oryzias latipes

“…In sea urchin, egg envelope hardening involves a multi-step process (1) including the formation of tyrosine-tyrosine cross-links between constituent proteins by ovoperoxidase (2, 8,14). On the other hand, y-glutamyl elysine cross-links were discovered in rainbow trout egg envelopes (chorions) hardened after fertilization (4), and a transglutaminase was suggested to participate in chorion hardening of cod eggs (15).…”