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Structure paper

TitleMolecular insights into capsular polysaccharide secretion.
Journal, issue, pagesNature, Year 2024
Publish dateApr 3, 2024
AuthorsJeremi Kuklewicz / Jochen Zimmer /
PubMed AbstractCapsular polysaccharides (CPSs) fortify the cell boundaries of many commensal and pathogenic bacteria. Through the ABC-transporter-dependent biosynthesis pathway, CPSs are synthesized intracellularly ...Capsular polysaccharides (CPSs) fortify the cell boundaries of many commensal and pathogenic bacteria. Through the ABC-transporter-dependent biosynthesis pathway, CPSs are synthesized intracellularly on a lipid anchor and secreted across the cell envelope by the KpsMT ABC transporter associated with the KpsE and KpsD subunits. Here we use structural and functional studies to uncover crucial steps of CPS secretion in Gram-negative bacteria. We show that KpsMT has broad substrate specificity and is sufficient for the translocation of CPSs across the inner bacterial membrane, and we determine the cell surface organization and localization of CPSs using super-resolution fluorescence microscopy. Cryo-electron microscopy analyses of the KpsMT-KpsE complex in six different states reveal a KpsE-encaged ABC transporter, rigid-body conformational rearrangements of KpsMT during ATP hydrolysis and recognition of a glycolipid inside a membrane-exposed electropositive canyon. In vivo CPS secretion assays underscore the functional importance of canyon-lining basic residues. Combined, our analyses suggest a molecular model of CPS secretion by ABC transporters.
External linksNature / PubMed:38570679
MethodsEM (single particle)
Resolution3.1 - 4.4 Å
Structure data

41592

8tsh

EMDB-41592, PDB-8tsh:
S. thermodepolymerans KpsMT(E151Q)-KpsE in complex with ATP
Method: EM (single particle) / Resolution: 3.1 Å

41593

8tsi

EMDB-41593, PDB-8tsi:
S. thermodepolymerans KpsMT-KpsE in complex with ADP:AlF4-
Method: EM (single particle) / Resolution: 4.4 Å

41595

8tsl

EMDB-41595, PDB-8tsl:
S. thermodepolymerans KpsM-KpsE in Apo 2 state with rigid body fitted KpsT
Method: EM (single particle) / Resolution: 3.4 Å

41601

8tsw

EMDB-41601, PDB-8tsw:
S. thermodepolymerans KpsMT-KpsE Apo 1
Method: EM (single particle) / Resolution: 3.1 Å

41602

8tt3

EMDB-41602, PDB-8tt3:
S. thermodepolymerans KpsM-KpsE in Glycolipid 2 state with rigid body fitted KpsT
Method: EM (single particle) / Resolution: 3.4 Å

41626

8tun

EMDB-41626, PDB-8tun:
S. thermodepolymerans KpsM-KpsE in Glycolipid 1 state with rigid body fitted KpsT
Method: EM (single particle) / Resolution: 3.4 Å

EMDB-41677: S. thermodepolymerans KpsMT-KpsE Apo 1 - consensus map
Method: EM (single particle) / Resolution: 3.1 Å

EMDB-41678: S. thermodepolymerans KpsMT-KpsE Apo 1 - KpsT-focused map
Method: EM (single particle) / Resolution: 3.3 Å

EMDB-41681: S. thermodepolymerans KpsMT(E151Q)-KpsE in complex with ATP - consensus map
Method: EM (single particle) / Resolution: 3.1 Å

EMDB-41682: S. thermodepolymerans KpsMT(E151Q)-KpsE in complex with ATP - crown focused map
Method: EM (single particle) / Resolution: 3.3 Å

EMDB-41697: S. thermodepolymerans KpsMT-KpsE in Apo 2 state - consensus map
Method: EM (single particle) / Resolution: 3.3 Å

EMDB-41698: S. thermodepolymerans KpsMT-KpsE in Apo 2 state - KpsT focused map
Method: EM (single particle) / Resolution: 3.5 Å

EMDB-41718: S. thermodepolymerans KpsMT-KpsE with bound glycolipid - state 1 - consensus map
Method: EM (single particle) / Resolution: 3.4 Å

EMDB-41720: S. thermodepolymerans KpsMT-KpsE with bound glycolipid - state 1 - KpsT focused map
Method: EM (single particle) / Resolution: 3.4 Å

EMDB-41721: S. thermodepolymerans KpsMT-KpsE with bound glycolipid - state 1 - KpsM focused map
Method: EM (single particle) / Resolution: 3.4 Å

EMDB-41771: S. thermodepolymerans KpsMT-KpsE with bound glycolipid - state 2 - consensus map
Method: EM (single particle) / Resolution: 3.4 Å

EMDB-41772: S. thermodepolymerans KpsMT-KpsE with bound glycolipid - state 2 - KpsT focused map
Method: EM (single particle) / Resolution: 3.4 Å

EMDB-41773: S. thermodepolymerans KpsMT-KpsE with bound glycolipid - state 2 - KpsM focused map
Method: EM (single particle) / Resolution: 3.4 Å

Chemicals

ChemComp-ATP:
ADENOSINE-5'-TRIPHOSPHATE / ATP, energy-carrying molecule*YM / Adenosine triphosphate

ChemComp-MG:
Unknown entry

ChemComp-ADP:
ADENOSINE-5'-DIPHOSPHATE / ADP, energy-carrying molecule*YM / Adenosine diphosphate

ChemComp-ALF:
TETRAFLUOROALUMINATE ION


ChemComp, No image

ChemComp-KJ9:
Unknown entry

Source
  • caldimonas thermodepolymerans (bacteria)
KeywordsMEMBRANE PROTEIN / ABC transporter / Capsular polysaccharide

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