Structure of a filamentous phosphoglycoprotein polymer

the secreted acid phosphatase of Leishmania mexicana

Y D Stierhof, M Wiese, T Ilg, P Overath, M Häner, U Aebi

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

The insect stage of the protozoan parasite Leishmania mexicana secretes a filamentous acid phosphatase (secreted acid phosphatase, SAP), a polymeric phosphoglycoprotein. The wild-type (wt) SAP filament is a copolymer composed of two related gene products SAP1 and SAP2, which are identical in the enzymatically active NH2-terminal domain and the COOH-terminal domain, but differ in the length of a highly glycosylated Ser/Thr-rich repeat region (32 amino acids and 383 amino acids, respectively) which is located between these domains. When expressed separately, full length SAP1, SAP2, or the NH2-terminal domain alone, are able to assemble into filaments. The Ser/Thr-rich region is the exclusive target for a novel type of O-glycosylation via phosphoserines. By using glycerol spraying/low-angle rotary metal shadowing and labelling with monoclonal antibodies it is demonstrated that the repetitive region adopts an extended conformation forming side arms which project radially from the filament core and terminate with the COOH-terminal domain. The length of the side arms of SAP1 and SAP2 (20 nm and 90 nm, respectively) corresponds to the predicted length of the Ser/Thr-rich repeat region of SAP1 and SAP2. Mass determination by scanning electron microscopy (STEM) shows that one morphologically defined globular particle of the filament core is a polypeptide dimer. We propose a model for the filament core, in which the globular NH2-terminal SAP domains form one strand composed of polypeptide dimers or two tightly associated strands of monomers which may twist into a double helix, similar to actin filaments. The highly O-glycosylated side arms project from the filament core conferring an overall bottle-brush-like appearance. The L. mexicana SAP is compared to SAPs secreted by the closely related species L. amazonensis and L. donovani.

Original languageEnglish
Pages (from-to)137-148
Number of pages12
JournalJournal of Molecular Biology
Volume282
Issue number1
DOIs
Publication statusPublished - 11 Sep 1998

Fingerprint

Leishmania mexicana
Acid Phosphatase
Polymers
Phosphoserine
Amino Acids
Peptides
Scanning Transmission Electron Microscopy
Nucleic Acid Repetitive Sequences
Actin Cytoskeleton
Glycosylation
Electron Scanning Microscopy
Glycerol
Insects
Parasites
Metals
Monoclonal Antibodies
Genes

Keywords

  • acid phosphatase
  • dimerization
  • gene deletion
  • glycoproteins
  • insects
  • Leishmania
  • Leishmania donovani
  • Leishmania mexicana
  • microscopy, electron, scanning transmission
  • models, molecular
  • mutagenesis
  • phosphoproteins
  • recombinant proteins
  • electron microscopy
  • filament structure
  • mass determination
  • O-glycosylation
  • scanning transmission electron microscopy

Cite this

Stierhof, Y D ; Wiese, M ; Ilg, T ; Overath, P ; Häner, M ; Aebi, U. / Structure of a filamentous phosphoglycoprotein polymer : the secreted acid phosphatase of Leishmania mexicana. In: Journal of Molecular Biology . 1998 ; Vol. 282, No. 1. pp. 137-148.
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Structure of a filamentous phosphoglycoprotein polymer : the secreted acid phosphatase of Leishmania mexicana. / Stierhof, Y D; Wiese, M; Ilg, T; Overath, P; Häner, M; Aebi, U.

In: Journal of Molecular Biology , Vol. 282, No. 1, 11.09.1998, p. 137-148.

Research output: Contribution to journalArticle

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T1 - Structure of a filamentous phosphoglycoprotein polymer

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AU - Stierhof, Y D

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