Identification of full-length dentin matrix protein 1 in dentin and bone.
Dentin matrix protein 1 (DMP1) has been identified in the extracellular matrix (ECM) of dentin and bone as the processed NH(2)-terminal and COOH-terminal fragment. However, the full-length form of DMP1 has not been identified in these tissues. The focus of this investigation was to search for the intact full-length DMP1 in dentin and bone. We used two types of anti-DMP1 antibodies to identify DMP1: one type specifically recognizes the NH(2)-terminal region and the other type is only reactive to the COOH-terminal region of the DMP1 amino acid sequence. An approximately 105-kDa protein, extracted from the ECM of rat dentin and bone, was recognized by both types of antibodies; and the migration rate of this protein was identical to the recombinant mouse full-length DMP1 made in eukaryotic cells. We concluded that this approximately 105-kDa protein is the full-length form of DMP1, which is considerably less abundant than its processed fragments in the ECM of dentin and bone. We also detected the full-length form of DMP1 and its processed fragments in the extract of dental pulp/odontoblast complex dissected from rat teeth. In addition, immunofluorescence analysis showed that in MC3T3-E1 cells the NH(2)-terminal and COOH-terminal fragments of DMP1 are distributed differently. Our findings indicate that the majority of DMP1 must be cleaved within the cells that synthesize it and that minor amounts of uncleaved DMP1 molecules are secreted into the ECM of dentin and bone.
Huang
Bingzhen
B
Department of Biomedical Sciences, Baylor College of Dentistry, Texas A & M University System Health Science Center, 3302 Gaston Avenue, Dallas, TX 75246, USA.
Maciejewska
Izabela
I
Sun
Yao
Y
Peng
Tao
T
Qin
Disheng
D
Lu
Yongbo
Y
Bonewald
Lynda
L
Butler
William T
WT
Feng
Jian
J
Qin
Chunlin
C
D016475:3T3 Cells; D000818:Animals; D001857:Bone Matrix / Q000737:chemistry*; D003782:Dental Pulp / Q000737:chemistry; D003804:Dentin / Q000737:chemistry* / Q000378:metabolism; D005109:Extracellular Matrix / Q000737:chemistry; D016326:Extracellular Matrix Proteins / Q000032:analysis* / Q000276:immunology; D005455:Fluorescent Antibody Technique; D006801:Humans; D007668:Kidney / Q000166:cytology / Q000378:metabolism; D051379:Mice; D010006:Osteoblasts / Q000166:cytology / Q000378:metabolism; D010446:Peptide Fragments / Q000737:chemistry; D010750:Phosphoproteins / Q000032:analysis* / Q000276:immunology; D011499:Protein Processing, Post-Translational; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry; D014162:Transfection
C081635:Dmp1 protein, rat; D016326:Extracellular Matrix Proteins; D010446:Peptide Fragments; D010750:Phosphoproteins; D011994:Recombinant Proteins