Effect of chicken lysozyme signal peptide alterations on secretion of human lysozyme in Saccharomyces cerevisiae.
To investigate the structural requirements and functions of the N-terminal and the C-terminal regions of the chicken lysozyme signal peptide, the amino acids of each region were altered. The replacement of Gly(-1) and Leu(-2) with Pro(-1) and Ala(-2) or Val(-2), respectively, resulted in the complete shift of the cleavage site from position -1 to -2 in yeast (Saccharomyces cerevisiae). This shows that the introduction of a turn-promoting residue like Pro makes it possible to control the cleavage site of the signal peptide. Deletion of the positive charge and introduction of a negative charge in the N-terminal region decreased the lytic activity of secreted human lysozyme (HLY) and processing efficiency of preHLY, but the length and additional positive charge in this region had little influence. This suggests that the length of the N-terminal region scarcely influences the function of the signal peptide and that this region possibly interacts with the endoplasmic reticulum membrane to initiate the translocation of preprotein, similar to prokaryotic signal peptide. However, it needs only minimum positive charge for its function.
Tsuchiya
Y
Y
Bioengineering Research Laboratory, National Institute of Animal Health, Ibaraki, Japan.
Fujisawa
H
H
Nakayama
K
K
Nagahora
H
H
Jigami
Y
Y
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002645:Chickens; D018014:Gene Transfer Techniques; D006801:Humans; D008969:Molecular Sequence Data; D009113:Muramidase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D010957:Plasmids; D021382:Protein Sorting Signals / Q000737:chemistry*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D013329:Structure-Activity Relationship
D021382:Protein Sorting Signals; D009113:Muramidase