Two novel targeting peptide degrading proteases, PrePs, in mitochondria and chloroplasts, so similar and still different
2005 (English)In: Journal of Molecular Biology, ISSN 0022-2836, Vol. 349, no 4, 847-860 p.Article in journal (Refereed) Published
Two novel metalloproteases from Arabidopsis thaliana, termed AtPrePI and AtPrePII, were recently identified and shown to degrade targeting peptides in mitochondria and chloroplasts using an ambiguous targeting peptide. AtPrePI and AtPrePII are classified as dually targeted proteins as they are targeted to both mitochondria and chloroplasts. Both proteases harbour an inverted metal binding motif and belong to the pitrilysin subfamily A. Here we have investigated the subsite specificity of AtPrePI and AtPrePII by studying their proteolytic activity against the mitochondrial F1β pre-sequence, peptides derived from the F1β pre-sequence as well as non-mitochondrial peptides and proteins. The degradation products were analysed, identified by MALDI-TOF spectrometry and superimposed on the 3D structure of the F 1β pre-sequence. AtPrePI and AtPrePII cleaved peptides that are in the range of 10 to 65 amino acid residues, whereas folded or longer unfolded peptides and small proteins were not degraded. Both proteases showed preference for basic amino acids in the P1 position and small, uncharged amino acids or serine residues in the P′1 position. Interestingly, both AtPrePI and AtPrePII cleaved almost exclusively towards the ends of the α-helical elements of the F1β pre-sequence. However, AtPrePI showed a preference for the N-terminal amphiphilic α-helix and positively charged amino acid residues and degraded the F1β pre-sequence into 10-16 amino acid fragments, whereas AtPrePII did not show any positional preference and degraded the F1β pre-sequence into 10-23 amino acid fragments. In conclusion, despite the high sequence identity between AtPrePI and AtPrePII and similarities in cleavage specificities, cleavage site recognition differs for both proteases and is context and structure dependent. © 2005 Elsevier Ltd. All rights reserved.
Place, publisher, year, edition, pages
2005. Vol. 349, no 4, 847-860 p.
National CategoryMedical and Health Sciences
IdentifiersURN: urn:nbn:se:liu:diva-30890DOI: 10.1016/j.jmb.2005.04.023Local ID: 16553OAI: oai:DiVA.org:liu-30890DiVA: diva2:251713