Innate immunity: Bacterial cell-wall muramyl peptide targets the conserved transcription factor YB-1.

The bacterial cell wall muramyl dipeptides MDP and glucosaminyl-MDP (GMDP) are powerful immunostimulators but their binding target remains controversial. We previously reported expression cloning of GMDP-binding polypeptides and identification of Y-box protein 1 (YB-1) as their sole target. Here we show specific binding of GMDP to recombinant YB-1 protein and subcellular colocalization of YB-1 and GMDP.

Search for ligand of N-acetylglucosaminyl-N-acetylmuramyl dipeptide using its peptide mimetic.

The method for searching for ligands exerting an adjuvant effect is described. The method involves isolation of polysomes using an immobilized peptide mimetic of N-acetylglucosaminyl-N-acetylmuramyl dipeptide (GMDP) - RN-peptide. After the affinity chromatography and washing, RN-peptide complexes with the target sequences were dissociated with guanidine hydrochloride.

Structural modification effects on bioactivities of the novel 15-mer peptide adjuvant.

Structure-function relationship studies for novel GMDP-peptide mimic, RVPPRYHAKISPMVN (RN-peptide) were performed on array of truncated and 'Ala-scan' analogues. The shortest peptide fragment possessing detectable affinity towards anti-GMDP-antibodies was demonstrated to be PRYH. RN-peptide analogues lacking up to 8 residues at C-terminus were found to retain adjuvant activity with the minimal active peptide being RVPPRYH.

Simultaneous detection of seven staphylococcal enterotoxins: development of hydrogel biochips for analytical and practical application.

A method of simultaneous analysis of staphylococcal enterotoxins using hydrogel-based microarrays (biochips) has been developed. The method allows simultaneous quantitative detection of seven enterotoxins: A, B, C1, D, E, G, and I in a single sample. The development of the method included expression and purification of recombinant toxins, production of panels of monoclonal antibodies (mAbs) against the toxins, and design and manufacturing of an experimental biochip for the screening of mAbs and selection of optimal pairs of primary and secondary antibodies for each toxin.

[A method for the preparation of adjuvant peptide mimetics of GMDP with the use of monoclonal antibodies and combinatorial libraries of peptides in the format of phage display].

A method for the preparation of peptide mimetics of GMDP which could exhibit adjuvant activity without the negative effects of GMDP is described. The search for peptides with GMDP-like adjuvant activity was performed using highly specific monoclonal antibodies against GMDP and combinatorial peptide libraries in the format of phage display. Various elution methods were used for the immunoaffinity enrichment of the libraries during the course of the preparation of highly active and specific peptides.