Protein N-homocysteinylation induces the formation of toxic amyloid-like protofibrils.
Previous works reported that a mild increase in homocysteine level is a risk factor for cardiovascular and neurodegenerative diseases in humans. Homocysteine thiolactone is a cyclic thioester, most of which is produced by an error-editing function of methionyl-tRNA synthetase, causing in vivo post-translational protein modifications by reacting with the epsilon-amino group of lysine residues. In cells, the rate of homocysteine thiolactone synthesis is strictly dependent on the levels of the precursor metabolite, homocysteine. In this work, using bovine serum albumin as a model, we investigated the impact of N-homocysteinylation on protein conformation as well as its cellular actions. Previous works demonstrated that protein N-homocysteinylation causes enzyme inactivation, protein aggregation, and precipitation. In addition, in the last few years, several pieces of evidence have indicated that protein unfolding and aggregation are crucial events leading to the formation of amyloid fibrils associated with a wide range of human pathologies. For the first time, our results reveal how the low level of protein N-homocysteinylation can induce mild conformational changes leading to the formation of native-like aggregates evolving over time, producing amyloid-like structures. Taking into account the fact that in humans about 70% of circulating homocysteine is N-linked to blood proteins such as serum albumin and hemoglobin, the results reported in this article could have pathophysiological relevance and could contribute to clarify the mechanisms underlying some pathological consequences described in patients affected by hyperhomocysteinemia.
Dipartimento di Scienze Biochimiche, Università degli Studi di Firenze, Viale Morgagni 50, 50134 Firenze, Italy. paolo.paoli@unifi.it
Paolo
P
Paoli
Francesca
F
Sbrana
Bruno
B
Tiribilli
Anna
A
Caselli
Barbara
B
Pantera
Paolo
P
Cirri
Alina
A
De Donatis
Lucia
L
Formigli
Daniele
D
Nosi
Giampaolo
G
Manao
Guido
G
Camici
Giampietro
G
Ramponi
D000682:Amyloid / Q000737:chemistry* / Q000378:metabolism*; D000818:Animals; D002417:Cattle; D006710:Homocysteine / Q000378:metabolism*; D006801:Humans; D046529:Microscopy, Electron, Transmission; D011487:Protein Conformation; D011499:Protein Processing, Post-Translational; D012710:Serum Albumin, Bovine / Q000737:chemistry* / Q000378:metabolism*
D000682:Amyloid; D006710:Homocysteine; D012710:Serum Albumin, Bovine