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TitleChitin Translocation Is Functionally Coupled with Synthesis in Chitin Synthase.
Journal, issue, pagesInt J Mol Sci, Vol. 25, Issue 21, Year 2024
Publish dateOct 30, 2024
AuthorsSuhao Niu / Lei Qi / Xiaoyue Zhang / Dongfang He / Pengwei Li / Hao Wang / Yunchen Bi /
PubMed AbstractChitin, an extracellular polysaccharide, is synthesized by membrane-embedded chitin synthase (CHS) utilizing intracellular substrates. The mechanism of the translocation of synthesized chitin across ...Chitin, an extracellular polysaccharide, is synthesized by membrane-embedded chitin synthase (CHS) utilizing intracellular substrates. The mechanism of the translocation of synthesized chitin across the membrane to extracellular locations remains unresolved. We prove that the chitin synthase from (CHS) is a processive glycosyltransferase, which can rapidly produce and tightly bind with the highly polymerized chitin. We further demonstrate that CHS is a bifunctional enzyme, which is necessary and sufficient to translocate the synthesized chitin. CHS was purified and then reconstituted into proteoliposomes (PLs). The nascent chitin is generated and protected from chitinase degradation unless detergent solubilizes the PLs, showing that CHS translocates the newly produced chitin into the lumen of the PLs. We also attempted to resolve the CHS structure of the synthesized chitin-bound state, although it was not successful; the obtained high-resolution structure of the UDP/Mn-bound state could still assist in describing the characterization of the CHS's transmembrane channel. Consistently, we demonstrate that CHS is indispensable and capable of translocating chitin in a process that is tightly coupled to chitin synthesis.
External linksInt J Mol Sci / PubMed:39519219 / PubMed Central
MethodsEM (single particle)
Resolution2.94 - 3.2 Å
Structure data

36893

8k52

EMDB-36893, PDB-8k52:
Cryo-EM structure of chitin synthase
Method: EM (single particle) / Resolution: 2.94 Å

39709

8z0o

EMDB-39709, PDB-8z0o:
Cryo-EM structure of chitin synthase
Method: EM (single particle) / Resolution: 3.2 Å

Chemicals

ChemComp-UDP:
URIDINE-5'-DIPHOSPHATE / UDP*YM

ChemComp-MN:
Unknown entry

Source
  • Phytophthora sojae (eukaryote)
  • phytophthora sojae strain p6497 (eukaryote)
KeywordsTRANSFERASE / chitin synthase / membrane-integrated glycosyltransferase

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