A probe or primer according to the disclosure preferably encompasses the nucleic acid codon which encodes the E533D variation within the GPR156 protein, or the complement thereof. In some embodiments, position 207 of this GPR156 protein (referring to SEQ ID NO:3) is a glutamic acid. For DNA-DNA hybrids, the TAlso provided are methods for detecting the presence or quantifying the levels of variant GPR156 polypeptide in a biological sample, including, for example, protein sequencing and immunoassays.
In some embodiments, the mRNA comprises or consists of a nucleic acid sequence that encodes a polypeptide having an amino acid sequence that has at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO:5, wherein the GPR156 protein comprises an aspartic acid at the position corresponding to position 529 according to SEQ ID NO:5.In some embodiments, the mRNA encodes the portion of the GPR156 protein corresponding to positions 310 to 814 of SEQ ID NO:4. A primer according to the disclosure may also be used to amplify a GPR156 isoform (such as the isoform having the amino acid sequence of SEQ ID NO:5) or a fragment thereof comprising the E529D mutation. In some embodiments, the detecting step comprises sequencing the entire nucleic acid molecule.In some embodiments, the detecting step comprises: amplifying at least a portion of the nucleic acid molecule that encodes a GPR156 protein, wherein the amplified nucleic acid molecule encodes an amino acid sequence which comprises the position corresponding to position 533 according to SEQ ID NO:4 (and/or to position 529 according to SEQ ID NO:5); labeling the nucleic acid molecule with a detectable label; contacting the labeled nucleic acid with a support comprising a probe, wherein the probe comprises a nucleic acid sequence which hybridizes under stringent conditions to a nucleic acid sequence encoding aspartic acid at the position corresponding to position at 533 according to SEQ ID NO:4 (and/or to position 529 according to SEQ ID NO:5); and detecting the detectable label. For example, linkages for amino acids or amino acid analogs include, but are not limited to, —CHIn some embodiments, the isolated polypeptides comprise D-amino acids, which can be used to generate more stable peptides because D amino acids are not recognized by peptidases. This cytosine corresponds to the third position of the codon encoding the aspartic acid at the position corresponding to position 533 according to SEQ ID NO:4).In some embodiments, the isolated nucleic acid molecules comprise a nucleic acid sequence comprising positions 1521 through 1680 of any of the cDNA or mRNA molecules disclosed herein, having a thymine at the position corresponding to position 1599.In some embodiments, the cDNA molecules comprise less than the entire cDNA sequence of GPR156. Thus, while each particular nucleic acid sequence may not be written out herein, each and every sequence is in fact disclosed and described herein through the disclosed polypeptide sequences.Percent identity (or percent complementarity) between particular stretches of nucleic acid sequences within nucleic acids or amino acid sequences within polypeptides can be determined routinely using BLAST programs (basic local alignment search tools) and PowerBLAST programs (Altschul et al., J. Mol.
In some embodiments, the cDNA molecules comprise or consist of at least about 5, at least about 8, at least about 10, at least about 12, at least about 15, at least about 20, at least about 25, at least about 30, at least about 35, at least about 40, at least about 45, at least about 50, at least about 60, at least about 70, at least about 80, at least about 90, at least about 100, at least about 200, at least about 300, at least about 400, at least about 500, at least about 600, at least about 700, at least about 800, at least about 900, at least about 1000, at least about 1100, at least about 1200, at least about 1300, at least about 1400, at least about 1500, at least about 1600, at least about 1700, at least about 1800, at least about 1900, at least about 2000, at least about 2100, at least about 2200, at least about 2300, or at least about 2400 contiguous nucleotides of SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, or SEQ ID NO:21.
The natural antioxidant system includes enzymatic (catalase, glutathione peroxidase, and superoxide dismutase (SOD)) and direct-acting nonenzymatic (ascorbic acid, polyphenols, lipoic acid, and carotenoids) compounds and indirectly acting antioxidants that form adducts with metals and thus reverse ROS generation [Anthocyanins (a Greek word; anthos means flower and kyanos means blue) are an especially interesting and vigorously studied naturally occurring plant compounds that belong to one of the six subgroups of widespread plant constituents known as flavonoids [In plants, the anthocyanin molecule occurs as a glycoside that contains sugar (glycone moiety, i.e., mostly glucose, or xylose, rhamnose, galactose, or arabinose) and a nonsugar component (aglycone, i.e., anthocyanidin).