High‐level expression and characterization of a glycosylated human cementum protein 1 with lectin activity

Abstract

This work aims to contribute to the knowledge of human cementum protein 1 (CEMP1), its conformational characteristics and influence during the biomineralization process. The results revealed that hrCEMP1 expressed in Pichia pastoris is a 2.4% glycosylated, thermostable protein which possesses a molecular mass of 28 770 Da. The circular dichroism spectrum indicated a secondary structure content of 28.6% of alpha-helix, 9.9% of beta-sheet and 61.5% of random-coil forms. Biological activity assays demonstrated that hrCEMP1 nucleates and regulates hydroxyapatite crystal growth. Hereby, it is demonstrated for the first time that CEMP1 has a (C-type) lectin-like activity and specifically recognizes mannopyranoside. The information produced by this biochemical and structural characterization may contribute to understand more fully the biological functions of CEMP1.

Analysis of Carbohydrates and Glycoconjugates by Matrix‐assisted Laser Desorption/Ionization Mass Spectrometry: An Update for 2015–2016

“…Thus, for example, the difference of 21.2 kDa between these two masses has been interpreted as showing that the glycan content of hen egg white ovomacroglobulin is 11.5% (Geng et al, 2015). Similarly differences between sequence and experimental masses of human cementum protein 1 have indicated 2.4% glycosylation (Romo‐Arévalo et al, 2016). Differences of about 160 mass units in the masses of peptide fragments from tarsal adhesive secretions from the desert locust ( Schistocerca gregaria ) and the Madagascar hissing cockroach ( Gromphadorhina portentosa ) have also been taken to imply glycosylation (Betz et al, 2016).…”

Cementum protein 1‐derived peptide (CEMP 1‐p1) modulates hydroxyapatite crystal formation in vitro

“…Cementum protein 1 (CEMP1) is a glycosylated, phosphorylated, and thermostable protein. Circular dichroism analyses revealed that its secondary structure is composed by 28.6% alpha‐helix, 9.9% of beta‐sheet, and 61.5% of random‐coil forms . CEMP1 is mainly expressed by cementoblasts, cementocytes, subpopulations of periodontal ligament cells, mesenchymal stem cells located paravascularly into the periodontal ligament, Hertwig's epithelial root sheath cells, and dental follicle cells .…”

“…Instead, many biologically active proteins are highly flexible or intrinsically disordered . CEMP1 may be considered as an intrinsically disordered protein (IDP), since its sequence contains high percentage of random coil secondary structures . Recently, we have demonstrated that CEMP1's N‐terminus ‐derived peptide promotes differentiation of periodontal ligament cells toward a “mineralizing‐like” phenotype, and more importantly, the bioactive peptide shows osteoinductive and osteogenic properties, which enhanced the physiological deposition and maturation of newly formed bone in critical‐sized calvarial defects in Wistar rats .…”

“…14 CEMP1 may be considered as an intrinsically disordered protein (IDP), since its sequence contains high percentage of random coil secondary structures. 1,2,13,15,16 Recently, we have demonstrated that CEMP1's N-terminus-derived peptide promotes differentiation of periodontal ligament cells toward a "mineralizing-like" phenotype, and more importantly, the bioactive peptide shows osteoinductive and osteogenic properties, which enhanced the physiological deposition and maturation of newly formed bone in critical-sized calvarial defects in Wistar rats. 17 However, the characterization of undefined protein regions anticipates the disclosure of innovative functions into current biological processes; therefore, this study was designed to show that CEMP1-p1 nucleates, has affinity, and promotes the growth of HA crystals in vitro.…”

Solution NMR structure of cementum protein 1 derived peptide (CEMP1‐p1) and its role in the mineralization process

“…9,10 Through the biochemical characterization of hrCEMP1, it was hypothesized that serine-rich regions could drive the HA crystal formation process and thus mimic the function of the entire protein during the mineralization process. 11 In this sense, a synthetic peptide CEMP1-p1 (MGTSSTDSQQAQHRRCSTSN) was designed with the first 20 amino acids present in the N-terminus of CEMP1. This peptide contains 25% serine and possesses intrinsic characteristics to stimulate HA crystal nucleation and growth.…”