title: Removal of acidic residues of the prodomain of PCSK9 increases its activity towards the LDL receptor.
abstract: Proprotein convertase subtilisin/kexin type 9 (PCSK9) binds to the low density lipoprotein receptor (LDLR) at the cell surface and mediates intracellular degradation of the LDLR. The amino-terminus of mature PCSK9, residues 31-53 of the prodomain, has an inhibitory effect on this function of PCSK9, but the underlying mechanism is not fully understood. In this study, we have identified two highly conserved negatively charged segments (residues 32-40 and 48-50, respectively) within this part of the prodomain and performed deletions and substitutions to study their importance for degradation of the LDLRs. Deletion of the acidic residues of the longest negatively charged segment increased PCSK9's ability to degrade the LDLR by 31%, whereas a modest 8% increase was observed when these residues were mutated to uncharged amino acids. Thus, both the length and the charge of this part of the prodomain were important for its inhibitory effect. Deletion of the residues of the shorter second negatively charged segment only increased PCSK9's activity by 8%. Substitution of the amino acids of both charged segments to uncharged residues increased PCSK9's activity by 36%. These findings indicate that the inhibitory effect of residues 31-53 of the prodomain is due to the negative charge of this segment. The underlying mechanism could involve the binding of this peptide segment to positively charged structures which are important for PCSK9's activity. One possible candidate could be the histidine-rich C-terminal domain of PCSK9.
authors.affiliation: Unit for Cardiac and Cardiovascular Genetics, Department of Medical Genetics, Oslo, Norway.
authors.forename: Øystein L
authors.initials: ØL
authors.lastname: Holla
authors.forename: Jon K
authors.initials: JK
authors.lastname: Laerdahl
authors.forename: Thea Bismo
authors.initials: TB
authors.lastname: Strøm
authors.forename: Kristian
authors.initials: K
authors.lastname: Tveten
authors.forename: Jamie
authors.initials: J
authors.lastname: Cameron
authors.forename: Knut Erik
authors.initials: KE
authors.lastname: Berge
authors.forename: Trond P
authors.initials: TP
authors.lastname: Leren
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution / Q000235:genetics; D000818:Animals; D002460:Cell Line; D017124:Conserved Sequence; D006801:Humans; D008969:Molecular Sequence Data; D000071449:Proprotein Convertase 9; D043484:Proprotein Convertases; D017434:Protein Structure, Tertiary; D011973:Receptors, LDL / Q000378:metabolism*; D017384:Sequence Deletion; D012697:Serine Endopeptidases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D011973:Receptors, LDL; C472125:PCSK9 protein, human; D000071449:Proprotein Convertase 9; D043484:Proprotein Convertases; D012697:Serine Endopeptidases