The Sec1-Munc18 protein VPS33B forms a uniquely bidirectional complex with VPS16B

J Biol Chem. 2023 Jun;299(6):104718. doi: 10.1016/j.jbc.2023.104718. Epub 2023 Apr 14.


Loss-of-function variants of vacuolar protein sorting proteins VPS33B and VPS16B (VIPAS39) are causative for arthrogryposis, renal dysfunction, and cholestasis syndrome, where early lethality of patients indicates that VPS33B and VPS16B play essential cellular roles. VPS33B is a member of the Sec1-Munc18 protein family and thought to facilitate vesicular fusion via interaction with soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complexes, like its paralog VPS33A in the homotypic fusion and vacuole sorting complex. VPS33B and VPS16B are known to associate, but little is known about the composition, structure, or function of the VPS33B-VPS16B complex. We show here that human VPS33B-VPS16B is a high molecular weight complex, which we expressed in yeast to perform structural, composition, and stability analysis. Circular dichroism data indicate VPS33B-VPS16B has a well-folded α-helical secondary structure, and size-exclusion chromatography-multiangle light scattering revealed a molecular weight of ∼315 kDa. Quantitative immunoblotting indicated a VPS33B:VPS16B ratio of 2:3. Expression of arthrogryposis, renal dysfunction, and cholestasis syndrome-causing VPS33B missense variants showed L30P disrupts complex formation but not S243F or H344D. Truncated VPS16B (amino acids 143 to 316) was sufficient to form a complex with VPS33B. Small-angle X-ray scattering and negative-staining EM revealed a two-lobed shape for VPS33B-VPS16B. Avidin tagging indicated that each lobe contains a VPS33B molecule, and they are oriented in opposite directions. We propose a structure for VPS33B-VPS16B that allows the VPS33B at each end to interact with separate SNARE bundles and/or SNAREpins, plus associated membrane components. These observations reveal the only known potentially bidirectional Sec1-Munc18 protein complex.

Keywords: ARC syndrome; Sec1–Munc18; VPS33B–VPS16B; bidirectional pentameric complex; intracellular trafficking; vesicle trafficking proteins.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Humans
  • Munc18 Proteins*
  • Renal Insufficiency*
  • SNARE Proteins / genetics
  • Syndrome
  • Vesicular Transport Proteins / genetics
  • Vesicular Transport Proteins / metabolism


  • Munc18 Proteins
  • SNARE Proteins
  • Vesicular Transport Proteins
  • VIPAS39 protein, human
  • VPS33B protein, human
  • STXBP1 protein, human

Supplementary concepts

  • Arthrogryposis renal dysfunction cholestasis syndrome

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