Dynamic processing of recombinant dentin sialoprotein-phosphophoryn protein.
Dentin sialoprotein (DSP) and phosphophoryn (PP) are the two noncollagenous proteins classically linked to dentin but more recently found in bone, kidney, and salivary glands. These two proteins are derived from a single copy DSP-PP gene. Although this suggests that the DSP-PP gene is first transcribed into DSP-PP mRNAs, which later undergo processing to yield the DSP and PP proteins, this mechanism has not yet been demonstrated because of the inability to identify a DSP-PP precursor protein from any cell or tissue sample. To study this problem, we utilized a baculovirus expression system to produce recombinant DSP-PP precursor proteins from a DSP-PP(240) cDNA, which represents one of several endogenous DSP-PP transcripts that influence various tooth mineralization phases. Our in vitro results demonstrate that DSP-PP(240) precursor proteins are produced by this system and are capable of self-processing to yield both DSP and PP proteins. We further demonstrated that purified recombinant DSP-PP(240), purified recombinant PP(240), and the native highly phosphorylated protein (equivalent to the PP(523) isoform) have proteolytic activity. These newly identified tissue proteases may play key roles in tissue modeling during organogenesis.
Godovikova
Valentina
V
Department of Cariology, Restorative Sciences and Endodontics, University of Michigan School of Dentistry, Ann Arbor, Michigan 48109-1078, USA.
Ritchie
Helena H
HH
D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D018388:Codon, Terminator; D018076:DNA, Complementary; D003804:Dentin / Q000737:chemistry*; D005091:Exons; D007438:Introns; D013058:Mass Spectrometry; D008957:Models, Genetic; D008969:Molecular Sequence Data; D010750:Phosphoproteins / Q000235:genetics* / Q000378:metabolism; D011498:Protein Precursors; D011499:Protein Processing, Post-Translational; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012795:Sialoglycoproteins / Q000235:genetics* / Q000378:metabolism*
D018388:Codon, Terminator; D018076:DNA, Complementary; D010750:Phosphoproteins; D011498:Protein Precursors; D011994:Recombinant Proteins; D012795:Sialoglycoproteins; C017630:phosphophoryn