Back to Search
Start Over
Comparative calcium binding of leucine-rich amelogenin peptide and full-length amelogenin
- Source :
- Le, TQ; Gochin, M; Featherstone, JDB; Li, W; & DenBesten, PK. (2006). Comparative calcium binding of leucine-rich amelogenin peptide and full-length amelogenin. European Journal of Oral Sciences, 114(SUPPL. 1), 320-326. doi: 10.1111/j.1600-0722.2006.00313.x. UCSF: Retrieved from: http://www.escholarship.org/uc/item/4w87s0sp, European journal of oral sciences, vol 114 Suppl 1, iss SUPPL. 1, European journal of oral sciences, vol 114 Suppl 1, iss s1
- Publication Year :
- 2006
- Publisher :
- eScholarship, University of California, 2006.
-
Abstract
- Leucine-rich amelogenin peptide (LRAP) is an alternately spliced amelogenin. LRAP is known to bind to hydroxyapatite, and has been shown to signal mesenchymal cells to proliferate, but its function in enamel formation is unclear. The purpose of this study was to determine the calcium-binding properties and structure of recombinant human LRAP (rLRAP) compared with full-length amelogenin (rH174). rLRAP and rH174 were synthesized in Escherichia coli and purified by affinity chromatography and reverse-phase high-performance liquid chromatography. Calcium binding was measured by isothermal titration calorimetry (ITC) at pH 7.5 and 25 degrees C, and raw data were analyzed by origin 7.0 software. The structure of rLRAP was analyzed by nuclear magnetic resonance (NMR) and circular dichroism (CD) in the absence or presence of Ca2+, pH 7.5 and 4.0, at 25 degrees C. Thermodynamic values showed that rLRAP had a Ca2+-binding affinity approximately 6.4-times greater than rH174. NMR and CD data revealed that rLRAP was randomly coiled, and that this structure was not altered by Ca2+, which bound to rLRAP and rH174 via ionic interactions. Unlike r174 (beta-spiral), rLRAP had a random-coiled structure. The calcium binding and structural differences between rLRAP and rH174 suggest that these proteins have different functions in enamel biomineralization.
- Subjects :
- Circular dichroism
Magnetic Resonance Spectroscopy
Protein Conformation
chemistry.chemical_element
amelogenins
Calcium
Calorimetry
enamel mineralization
Chromatography, Affinity
Structure-Activity Relationship
Protein structure
Affinity chromatography
Dental Enamel Proteins
Amelogenesis
Escherichia coli
Humans
Dental/Oral and Craniofacial Disease
Dental Enamel
General Dentistry
Chromatography, High Pressure Liquid
Chromatography
Amelogenin
Chemistry
Circular Dichroism
Titrimetry
Temperature
Isothermal titration calorimetry
Nuclear magnetic resonance spectroscopy
Hydrogen-Ion Concentration
Recombinant Proteins
thermodynamic values
Enamel mineralization
Durapatite
Biochemistry
Affinity
High Pressure Liquid
Dentistry
Thermodynamics
leucine-rich amelogenin peptide
Protein Binding
Subjects
Details
- Language :
- English
- Database :
- OpenAIRE
- Journal :
- Le, TQ; Gochin, M; Featherstone, JDB; Li, W; & DenBesten, PK. (2006). Comparative calcium binding of leucine-rich amelogenin peptide and full-length amelogenin. European Journal of Oral Sciences, 114(SUPPL. 1), 320-326. doi: 10.1111/j.1600-0722.2006.00313.x. UCSF: Retrieved from: http://www.escholarship.org/uc/item/4w87s0sp, European journal of oral sciences, vol 114 Suppl 1, iss SUPPL. 1, European journal of oral sciences, vol 114 Suppl 1, iss s1
- Accession number :
- edsair.doi.dedup.....aece8f4c81fce08dc477554420b27f0e