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J. Bacteriol., Apr 1995, 1703-1711, Vol 177, No. 7
JW Sun, SY Wanda and R Curtiss 3rd
The dextranase inhibitor gene (dei) from Streptococcus sobrinus UAB108 was
previously cloned, expressed, and sequenced. Its gene product (Dei) has now
been purified as a single band with apparent molecular mass of 43 kDa, as
determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
The specific activity of Dei increased 121-fold upon purification. Most Dei
activity (91.2%) was located in the periplasmic fraction from recombinant
Escherichia coli cells. Dei competitively inhibits dextranase (Dex). This
competitive inhibition mechanism has been further shown by detection and
recovery of the intermediate enzyme- inhibitor (Dex-Dei) complex by gel
filtration technology using fast protein liquid chromatography. Calibration
of their molecular masses indicated that native Dei exists as a tetramer,
Dex exists as dimer, and the Dex-Dei complex consists of two Dex molecules
with two Dei molecules. Deletion analysis indicates that the intact Dei
molecule is essential for Dei activity but not for glucan binding and
immune cross- reaction. Dei is a special kind of glucan-binding protein
with ability to inhibit Dex with high specificity. It can inhibit
endogenous Dex, which can make more branches in glucan with the cooperation
of the glucosyltransferase GTF-I. This inhibition cause the accumulation of
water-soluble glucan. The latter reaction product can inhibit plaque
formation and adherence of the mutans group of streptococcal cells. Dei
derived from S. sobrinus UAB108 can inhibit only Dex from S. sobrinus
(serotypes d and g), S. downei (previously S. sobrinus, serotype h), and S.
macacae (serotype h).(ABSTRACT TRUNCATED AT 250 WORDS)
Copyright © 1995, American Society for Microbiology
Purification, characterization, and specificity of dextranase inhibitor (Dei) expressed from Streptococcus sobrinus UAB108 gene cloned in Escherichia coli [published erratum appears in J Bacteriol 1997 Jan;179(2):568]
Department of Biology, Washington University, St. Louis, Missouri 63130, USA.
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