Derived protein and cDNA sequences of hamster amelogenin.
Hamster enamel protein extracts were analyzed by RP-HPLC and the isolated fractions by SDS-and Western blotting using polyclonal antibodies against recombinant mouse amelogenin and anti-peptide antibodies against the mouse exon 4-encoded sequence. Total RNA was extracted from enamel organ epithelia and, using a 3' rapid amplification of cDNA ends (3' RACE) technique, the coding regions for three different amelogenin isoforms were cloned along with the 3' non-coding region. DNA sequencing revealed that the hamster amelogenin isoforms are 180, 73 and 59 amino acids in length, respectively. The 59-residue amelogenin corresponds to the leucine-rich amelogenin protein (LRAP), the 73-residue amelogenin corresponds to LRAP with the inclusion of the exon 4-encoded sequence, while the 180-residue amelogenin is the most abundant amelogenin isoform. Edman degradation was performed on purified hamster amelogenin, which provided the amino acid sequence in the region encoded by the 5' PCR amplification primer used in cloning. Therefore, the entire derived amino acid sequence of hamster amelogenin was revealed. The hamster amelogenin amino acid sequence was aligned with all its known homologues. Hamster differs from rat and mouse amelogenin at only three amino acid positions. Southern blot analysis using a panel of restriction enzymes gave the same pattern for hamster DNA obtained from males and females, suggesting that in hamster, as in mouse, amelogenin is expressed from a single gene located on the X chromosome.
Lyaruu
D M
DM
Tooth Dev. Sect., Dept. Oral Cell Biol., ACTA, Vrije Universiteit, Amsterdam, The Netherlands. dm.lyaruu.ocb.acta@med.vu.nl
Hu
C C
CC
Zhang
C
C
Qian
Q
Q
Ryu
O H
OH
Moradian-Oldak
J
J
Wöltgens
J H
JH
Fincham
A G
AG
Simmer
J P
JP
D053523:Amelogenin; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001483:Base Sequence; D006224:Cricetinae; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics*; D003746:Dental Enamel Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D007120:Immunochemistry; D051379:Mice; D008969:Molecular Sequence Data; D009805:Odontogenesis / Q000235:genetics; D016133:Polymerase Chain Reaction; D051381:Rats; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D014960:X Chromosome / Q000235:genetics
D053523:Amelogenin; C507200:Amelx protein, mouse; C507201:Amelx protein, rat; D000596:Amino Acids; D017931:DNA Primers; D018076:DNA, Complementary; D003746:Dental Enamel Proteins; C067432:leucine-rich amelogenin peptide