TY  - JOUR
N1  - Copyright of this article belongs to Wiley Online
ID  - open1758
UR  - http://dx.doi.org/10.1002/prot.24865
IS  - 10
A1  - Sebastian Samuel, Jesse
A1  - Kumar, Deepak
A1  - Babu Chodisetti, Sathi
A1  - Agrewala, Javed N
A1  - Singh, Balvinder
A1  - Guptasarma, Purnananda
A1  - Sarkar, Dibyendu
Y1  - 2015/10//
N2  - Although unglycosylated HuEpo is fully functional, it has very short serum half-life. However, the mechanism of in vivo clearance of human Epo (HuEpo) remains largely unknown. In this study, the relative importance of protease-sensitive sites of recombinant HuEpo (rHuEpo) has been investigated by analysis of structural data coupled with in vivo half-life measurements. Our results identify ?3-?4 inter-helical loop region as a target site of lysosomal protease Cathepsin L. Consistent with previously-reported lysosomal degradation of HuEpo, these results for the first time identify cleavage sites of rHuEpo by specific lysosomal proteases. Furthermore, in agreement with the lowered exposure of the peptide backbone around the cleavage site, remarkably substitutions of residues with bulkier amino acids result in significantly improved in vivo stability. Together, these results have implications for the mechanism of in vivo clearance of the protein in humans.
PB  - Wiley Online
JF  - Proteins
VL  - 83
KW  - cathepsin L; circulating half-life; in vivo clearance; inter-helical loop region; proteolysis; recombinant HuEpo
SN  - 1097-0134
TI  - Probing protease sensitivity of recombinant human erythropoietin reveals ?3-?4 inter-helical loop as a stability determinant.
SP  - 1813
EP  - 22
ER  -