Cianci, M. | Search bioRxiv

bioRxiv2026

Structural basis of the lobster carapace blue colour mediated by an HPR protein

The chemical basis underlying the striking blue hue of live H. americanus, known as American lobster, are studied in evolutionary biology and in polyene physical chemistry. Carapace colouration is generated by the antioxidant astaxanthin bound within the carotenoprotein crustacyanin complexes. Here, we present the ex vivo structure of the most abundant -crustacyanin and {beta}-crustacyanin forms, determined respectively by cryo-electron microscopy and X-ray crystallography to a resolution of 2.75 [A]. Our structural analysis reveals -crustacyanin as an elongated arrangement of {beta}-crustacyanin heterodimers tethered by an heptatricopeptide repeat (HPR) protein. In vitro complex formation between the {beta}-crustacyanin unit with a synthetic heptatricopeptide reproduces the observed blue colour of -crustacyanin, identifying the HPR protein, in concert with crustacyanins, as contributor in tuning carapace colour. Overall, these results explain how nature adjusts the colour across the entire visible spectrum by exploiting the bathochromic shift of astaxanthin from its unbound red form ({lambda}max = 472 nm) firstly to the {beta}-crustacyanin violet bound form ({lambda}max = 591 nm), and then to the -crustacyanin bound blue form ({lambda}max = 631 nm).

biochemistry↗

bioRxiv2023

3-O-methyltolcapone and Its Lipophilic Analogues are Potent Inhibitors of Transthyretin Amyloidogenesis with High Permeability and Low Toxicity

Transthyretin (TTR) is an amyloidogenic homotetramer involved in the transport of thyroxine in blood and cerebrospinal fluid. To date, more than 130 TTR point mutations are known to destabilise the TTR tetramer, leading to its extracellular pathological aggregation accumulating in several organs, such as heart, peripheral and autonomic nerves, and leptomeninges. Tolcapone is an FDA-approved drug for Parkinsons disease that has been repurposed as a TTR stabiliser. We characterised 3-O-methyltolcapone and two newly synthesized lipophilic analogues, which are expected to be protected from the metabolic glucuronidation which is responsible of the lability of tolcapone in the organism. Immunoblotting assays indicated the high degree of TTR stabilisation, coupled with binding selectivity towards TTR in diluted plasma of 3-O-methyltolcapone and its lipophilic analogues. Furthermore, in-vitro toxicity data showed their several-fold improved neuronal and hepatic safety compared to tolcapone. Calorimetric and structural data showed that both T4 binding sites of TTR are occupied by 3-O-methyltolcapone and its lipophilic analogs, consistent with an effective TTR tetramer stabilisation. Moreover, in-vitro permeability studies showed that the three compounds can effectively cross the blood-brain barrier, which is a prerequisite for the inhibition of TTR amyloidogenesis in the cerebrospinal fluid. Our data demonstrate the relevance of 3-O-methyltolcapone and its lipophilic analogs as potent inhibitors of TTR amyloidogenesis.

biochemistry↗