Linked Life Data

8600179

Source:http://linkedlifedata.com/resource/pubmed/id/8600179

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
12
pubmed:dateCreated
1996-4-26
pubmed:abstractText
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 final product as re-Hst1. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) showed that native histatin 1 and re-Hst1 had the same apparent molecular weights, while cationic PAGE showed that re-Hst1 was more basic. Phosphate analysis indicated 1 mol phosphate/mol of native histatin 1, while re-Hst1 lacked any detectable phosphate. Re-Hst1 demonstrated candidacidal activity comparable to that of native histatin 1, but displayed substantially lower binding to hydroxyapatite. These results show that phosphorylation of histatin 1 at residue 2 contributes significantly to its ability to bind to hydroxyapatite.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
D
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Dec
pubmed:issn
0022-0345
pubmed:author
pubmed:issnType
Print
pubmed:volume
74
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
1837-44
pubmed:dateRevised
2008-11-21
pubmed:meshHeading
pubmed-meshheading:8600179-Antifungal Agents, pubmed-meshheading:8600179-Base Sequence, pubmed-meshheading:8600179-Candida albicans, pubmed-meshheading:8600179-Cloning, Molecular, pubmed-meshheading:8600179-DNA Primers, pubmed-meshheading:8600179-Durapatite, pubmed-meshheading:8600179-Escherichia coli, pubmed-meshheading:8600179-Histatins, pubmed-meshheading:8600179-Humans, pubmed-meshheading:8600179-Molecular Sequence Data, pubmed-meshheading:8600179-Phosphopeptides, pubmed-meshheading:8600179-Phosphorylation, pubmed-meshheading:8600179-Phosphoserine, pubmed-meshheading:8600179-Protein Binding, pubmed-meshheading:8600179-Protein Structure, Secondary, pubmed-meshheading:8600179-Recombinant Fusion Proteins, pubmed-meshheading:8600179-Salivary Proteins and Peptides, pubmed-meshheading:8600179-Structure-Activity Relationship
pubmed:year
1995
pubmed:articleTitle
Functional comparison of native and recombinant human salivary histatin 1.
pubmed:affiliation
Department of Periodontology and Oral Biology, School of Graduate Dentistry, Boston University Medical Center, MA 02118, USA.
pubmed:publicationType
Journal Article, Comparative Study, Research Support, U.S. Gov't, P.H.S.