PMID: 8600179Dec 1, 1995Paper

Functional comparison of native and recombinant human salivary histatin 1

Journal of Dental Research
J DriscollFrank G Oppenheim

Abstract

Histatin 1 is a histidine-rich phosphoprotein present in human parotid saliva that possesses candidacidal activity and functions in mineralization by adsorbing to hydroxyapatite. The objective of the present study was to develop a system for recombinant production of histatin 1 and to examine the role of phosphorylation in the functional activities of this molecule. Native histatin 1 (containing a phosphoserine at residue 2) was purified from parotid saliva, whereas a bacterial expression system was used to produce a recombinant form of histatin 1 (re-Hst1) that lacked phosphorylated serine. Histatin 1 cDNA was inserted into the vector pGEX-3X, which expresses foreign genes as soluble fusion proteins attached to the carboxyl-terminus of glutathione S-transferase (GST). The GST/re-Hst1 fusion protein was isolated from cell lysates by affinity chromatography on glutathione (GSH)-Sepharose and digested with cyanogen bromide to separate re-Hst1 from the GST fusion partner. The digest was subjected to reversed-phase high-performance liquid chromatography on a C18 column, and re-Hst1 was eluted as a well-defined peak. The yield of re-Hst1 was 4 mg/L of bacterial culture. Amino-terminal sequencing and amino acid analysis confirmed the...Continue Reading

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Citations

May 20, 1999·Biopolymers·D Andreu, L Rivas
Jul 25, 2007·Antimicrobial Agents and Chemotherapy·Akira ShimotoyodomeYoshinori Takaesu
Mar 31, 2010·Journal of Dental Research·W L SiqueiraF G Oppenheim
Mar 18, 2004·The Journal of Pharmacy and Pharmacology·Kevin Kavanagh, Susan Dowd
Nov 24, 1998·Critical Reviews in Oral Biology and Medicine : an Official Publication of the American Association of Oral Biologists·H Tsai, L A Bobek
Jul 27, 2007·Journal of Dental Research·E J Helmerhorst, F G Oppenheim
Jun 28, 2015·Journal of Dental Research·Y XiaoW L Siqueira
Nov 16, 2010·Journal of Dental Research·E E McDonaldW L Siqueira
Nov 15, 2001·Journal of Dental Research·M S Lamkin, P Lindhe
Dec 13, 2017·Oral Diseases·P TorresV A Torres
Jun 1, 2011·Probiotics and Antimicrobial Proteins·Adam Pepperney, Michael L Chikindas
Dec 1, 1996·Infection and Immunity·H TsaiL A Bobek

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