1887

Abstract

The recombinant primer-dependent glucosyltransferase GtfJ of possesses a C-terminal glucan-binding domain composed of eighteen 21 aa YG repeats. By engineering a series of C-terminal truncated proteins, the position at which truncation prevented further mutan synthesis was defined to a region of 43 aa, confirming that not all of the YG motifs were required for the formation of mutan by GtfJ. The role of the YG repeats in glucan binding was investigated in detail. Three proteins consisting of 38, 72 or 110 C-terminal YG repeats were expressed in . Each of the three purified proteins bound to both the 1,6-α-linked glucose residues of dextran and the 1,3-α-linked glucose residues of mutan, indicating that a protein consisting of nothing but 38 YG repeats could attach to either substrate. Secondary structure predictions of the primary amino acid sequence suggested that 37% of the amino acids were capable of forming a structure such that five regions of β-sheet were separated by regions capable of forming β-turns and random coils. CD spectral analysis showed that the purified 38 YG protein possessed an unordered secondary structure with some evidence of possible β-sheet formation and that the protein maintained this relatively unordered structure on binding to dextran.

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2002-02-01
2019-10-19
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