Analysis of amelogenin proteins using monospecific antibodies to defined sequences

被引:19
作者
Gibson, CW
Kucich, U
Collier, P
Shen, G
Decker, S
Bashir, M
Rosenbloom, J
机构
[1] UNIV PENN,SCH DENT MED,ORAL BIOL RES CTR,PHILADELPHIA,PA 19104
[2] UNIV PENN,SCH DENT MED,DEPT ANAT & HISTOL,PHILADELPHIA,PA 19104
关键词
amelogenins; enamel; monospecific antibodies; alternative splicing;
D O I
10.3109/03008209509013711
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Amelogenins are the predominant proteins found in the developing enamel matrix and are believed to play a crucial role in normal mineralization. Although the amelogenin gene is found as a single copy in all species in which it has been examined, multiple amelogenin polypeptides ranging in size from 5 to 25 kDa are obtained upon extraction of developing enamel matrix, making identification and characterization of individual components difficult. This heterogeneity may be ascribed to transcription of divergent genes located on the X and Y chromosomes, alternative splicing of the primary transcripts, physiologic degradative processing, and artefactual degradation. Zn order to characterize individual components, antibodies were produced to the following peptides: (1) QPLQPMQPMQPLQPLQPL (corresponding to the repeat sequence encoded only in the bovine X chromosome gene), (2) IRHPPLPP (corresponding to a unique sequence generated by alternative splicing found in leucine-rich amelogenin peptide (LRAP), (3) LPDLPLEAWPATDKTKREEVD corresponding to the amelogenin carboxy-terminus. Amelogenin proteins obtained from fetal bovine molars were subjected to SDS PAGE and Western electrotransfer, and immuno-ultrastructural analysis. These analyses demonstrated that: (1) the distribution of amelogenin polypeptides isolated from male fetuses differed appreciably from that of females, (2) the LRAP junctional peptide sequence can be specifically identified, and (3) the LRAP peptide can be immunolocalized in the enamel matrix of both males and females.
引用
收藏
页码:109 / 114
页数:6
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