Water-soluble polypeptides comprised of repeat modules, method for preparing the same and method for a target-specific polypeptide and analysis of biological activity thereof
The present invention relates to a soluble polypeptide comprised of repeat modules. More particularly, the present invention relates to a soluble fusion polypeptide of the N-terminal domain of internalin and LRR (Leucine rich repeat) family protein, a method for preparing the polypeptide, a vector comprising a nucleic acid sequence encoding the polypeptide, a host cell comprising the vector, a method for producing a solubility and folding-improved fusion polypeptide by expressing the vector in the host cell, and a method for improving the solubility and folding of the fusion polypeptide. Further, the present invention relates to a method for preparing the polypeptide bound with a specific target and analyzing the efficacy of the soluble polypeptide.
1. A water-soluble fusion polypeptide prepared by fusion of the N-terminal domain of internalin protein, and LRR (Leucine-rich repeat) family protein,
wherein the fusion polypeptide has an amino acid sequence selected from the group consisting of SEQ ID NO:7, 8, 9, 10, 12, 15, 16, 17 and 18.
2. A method for analyzing the efficacy of the soluble fusion polypeptide of claim 1 , comprising the steps of:
(a) differentiating THP-1 cell line using PMA (phorbol 12-myristate 13-acetate);
(b) mixing in advance MD-2 (Myeloid differentiation protein-2), LPS (lipopolysaccharide), and the soluble fusion polypeptides of claim 1 so as to prepare mixtures;
(c) contacting the cell line of step (a) with the prepared mixtures of step (b); and
(d) determining that the soluble fusion polypeptide of claim 1 of the experimental group has better efficacy than that of the control group, when the experimental group contacted with the soluble fusion polypeptide of claim 1 shows a lower TNF-α (tumor necrosis factor alpha) level than the control group not contacted with the soluble fusion polypeptide of claim 1 to be analyzed.