Enamel matrix serine proteinase 1: stage-specific expression and molecular modeling.
Enamel proteins are cleaved by proteinases soon after their secretion by ameloblasts. Intact proteins concentrate in the outer enamel at or near the growing tips of the enamel crystallites while cleavage products accumulate in the deeper enamel. In the transition and early maturation stages there is a dramatic increase in proteolytic activity. This activity, coupled with the diminished secretory and increased reabsorptive functions of ameloblasts, leads to a precipitous fall in the amount of enamel protein in the matrix. Recently we have cloned and characterized an mRNA encoding a tooth-specific serine proteinase designated enamel matrix serine proteinase 1 (EMSP1) [Simmer et al., JDR (1998) 77: 377]. EMSP1 can be detected in the inner enamel during the secretory stage and its activity increases sharply during the transition stage. Stage-specific Northern blot analysis demonstrates this increase is accompanied by a parallel increase in the amount EMSP1 mRNA. A 3-dimensional computer model of EMSP1, based upon the crystal structure of bovine trypsin, has been generated and analyzed. All six disulfide bridges as well as the active site are conserved. Changes in the peptide binding region and the specificity pocket suggest that interaction of the proteinase with protein substrates is altered, potentially causing a shift in substrate specificity. The calcium binding region of trypsin is thoroughly modified suggesting that the calcium independence of EMSP1 activity is due to an inability to bind calcium. The three potential N-linked glycosylation sites, N104, N139 and N184, are in surface accessible positions away from the active site.
Scully
J L
JL
Department of Chemistry, Greenhills, Ann Arbor, MI, USA.
Bartlett
J D
JD
Chaparian
M G
MG
Fukae
M
M
Uchida
T
T
Xue
J
J
Hu
C C
CC
Simmer
J P
JP
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern / Q000379:methods; D002417:Cattle; D018076:DNA, Complementary; D004658:Enamel Organ / Q000254:growth & development; D018507:Gene Expression Regulation, Developmental; D007610:Kallikreins; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D012333:RNA, Messenger / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D012697:Serine Endopeptidases / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D013552:Swine; D014357:Trypsin / Q000737:chemistry
D018076:DNA, Complementary; D012333:RNA, Messenger; D007610:Kallikreins; D012697:Serine Endopeptidases; C120801:kallikrein 4; D014357:Trypsin