Synthesis and structural characterization of the peptide epitope of the ovarian cancer biomarker CA125 (MUC16).
A highly conserved region of 21 amino acids flanked by cysteine residues, contained within a larger repeated domain, has been proposed to be the antibody-binding site in the ovarian cancer biomarker CA125 (MUC16). In this study solid-phase peptide synthesis with Fmoc protection chemistry was used to assemble a 21-mer peptide corresponding to the most frequently occurring antibody binding sequence in CA125. Potentially significant sequence variants were also synthesized. Peptide secondary structure was investigated using Fourier transform infrared spectroscopy, revealing the consensus sequence peptide to be largely unstructured at physiological pH whether the cysteine residues were reduced or were oxidized to form an intramolecular disulfide bond. Substitution of serine for proline at position 8 (P8S) results in β-sheet formation in peptides involved in intramolecular disulfide bonds. This β-sheet structure does not persist in peptides incapable of intramolecular disulfide bonding because of sequence nor in peptides treated with the reducing agent dithiothreitol. In CA125, P8S is predicted to occur in ∼25% of repeat domains, suggesting that this structural motif is a non-negligible contributor to overall structure and function. These findings suggest that future structural characterization efforts of CA125 should be especially mindful of the amino acid sequence and oxidation state of the protein.
Berman
Zach T
ZT
Department of Chemistry and Biochemistry, Oberlin College, 119 Woodland Street, Oberlin, OH 44074, USA.
Moore
Lee J
LJ
Knudson
Kathleen E
KE
Whelan
Rebecca J
RJ
D000595:Amino Acid Sequence; D014408:Biomarkers, Tumor / Q000138:chemical synthesis* / Q000737:chemistry*; D018394:CA-125 Antigen / Q000737:chemistry*; D018985:Epitopes, B-Lymphocyte / Q000737:chemistry*; D005260:Female; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry*; D010051:Ovarian Neoplasms / Q000378:metabolism; D017433:Protein Structure, Secondary; D017550:Spectroscopy, Fourier Transform Infrared
D014408:Biomarkers, Tumor; D018394:CA-125 Antigen; D018985:Epitopes, B-Lymphocyte; C451759:MUC16 protein, human; D008565:Membrane Proteins