Expression and species-specific glycosylation of Leishmania mexicana secreted acid phosphatase in Leishmania major

M Wiese, I Görcke, P Overath

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Parasitic protozoa of the genus Leishmania cycle between two main developmental stages, the promastigote in the digestive tract of the sandfly and the amastigote in the mononuclear cells of the mammalian host. Both life stages have been shown to secrete a number of highly glycosylated molecules [1] and [2]. The SAP of Leishmania mexicana, a component found in large amounts in the culture supernatant of promastigotes, is the best studied example of this group of proteins. It is a copolymer of 100 kDa (SAP1) and 200 kDa (SAP2) subunits which assemble into long filaments [3]. The backbone of the filaments is formed by a ‘string-of-pearls-like’ arrangement of globular protein subunits [4] and [5]. Projecting from this filament, two types of side arms differing in length have been demonstrated by glycerol spraying/low-angle rotary metal shadowing conferring an overall ‘bottle-brush-like’ appearance [5]. These side arms correspond to the Ser/Thr-rich repeat regions of 32 and 383 amino acids of SAP1 and SAP2, respectively, which have been shown to be the exclusive targets for glycosylation via phosphoserines, and a conserved COOH-terminal domain of 54 amino acids [3] and [6]. The composition of the phosphoserine-linked glycans has been studied in detail using monoclonal antibodies directed against phosphodi- and phosphotrisaccharide subunits and oligomannose cap structures [7] and chemical structure analysis [6]. These and other studies showed that SAP shares carbohydrate epitopes with LPG, the major glycolipid component of the promastigote surface [6], [7], [8] and [9]. Here, we show that the Ser/Thr-rich repetitive region of the secreted acid phosphatase of L. mexicana serves as a target for species-specific phosphoglycosylation in Leishmania major.
Original languageEnglish
Pages (from-to)325-329
Number of pages5
JournalMolecular and Biochemical Parasitology
Volume102
Issue number2
DOIs
Publication statusPublished - 20 Aug 1999

Fingerprint

Leishmania mexicana
Phosphoserine
Leishmania major
Acid Phosphatase
Glycosylation
Penicillin G Benzathine
Amino Acids
Psychodidae
Nucleic Acid Repetitive Sequences
Leishmania
Glycolipids
Protein Subunits
Glycerol
Polysaccharides
Gastrointestinal Tract
Epitopes
Metals
Monoclonal Antibodies
Carbohydrates
Proteins

Keywords

  • acid phosphatase
  • fluorescent antibody technique
  • glycosylation
  • immunoblotting
  • Leishmania major
  • Leishmania mexicana
  • plasmids
  • recombinant proteins
  • species specificity
  • secreted acid phosphatase
  • phosphoglycan modification
  • filament formation

Cite this

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title = "Expression and species-specific glycosylation of Leishmania mexicana secreted acid phosphatase in Leishmania major",
abstract = "Parasitic protozoa of the genus Leishmania cycle between two main developmental stages, the promastigote in the digestive tract of the sandfly and the amastigote in the mononuclear cells of the mammalian host. Both life stages have been shown to secrete a number of highly glycosylated molecules [1] and [2]. The SAP of Leishmania mexicana, a component found in large amounts in the culture supernatant of promastigotes, is the best studied example of this group of proteins. It is a copolymer of 100 kDa (SAP1) and 200 kDa (SAP2) subunits which assemble into long filaments [3]. The backbone of the filaments is formed by a ‘string-of-pearls-like’ arrangement of globular protein subunits [4] and [5]. Projecting from this filament, two types of side arms differing in length have been demonstrated by glycerol spraying/low-angle rotary metal shadowing conferring an overall ‘bottle-brush-like’ appearance [5]. These side arms correspond to the Ser/Thr-rich repeat regions of 32 and 383 amino acids of SAP1 and SAP2, respectively, which have been shown to be the exclusive targets for glycosylation via phosphoserines, and a conserved COOH-terminal domain of 54 amino acids [3] and [6]. The composition of the phosphoserine-linked glycans has been studied in detail using monoclonal antibodies directed against phosphodi- and phosphotrisaccharide subunits and oligomannose cap structures [7] and chemical structure analysis [6]. These and other studies showed that SAP shares carbohydrate epitopes with LPG, the major glycolipid component of the promastigote surface [6], [7], [8] and [9]. Here, we show that the Ser/Thr-rich repetitive region of the secreted acid phosphatase of L. mexicana serves as a target for species-specific phosphoglycosylation in Leishmania major.",
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Expression and species-specific glycosylation of Leishmania mexicana secreted acid phosphatase in Leishmania major. / Wiese, M; Görcke, I; Overath, P.

In: Molecular and Biochemical Parasitology, Vol. 102, No. 2, 20.08.1999, p. 325-329.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Expression and species-specific glycosylation of Leishmania mexicana secreted acid phosphatase in Leishmania major

AU - Wiese, M

AU - Görcke, I

AU - Overath, P

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N2 - Parasitic protozoa of the genus Leishmania cycle between two main developmental stages, the promastigote in the digestive tract of the sandfly and the amastigote in the mononuclear cells of the mammalian host. Both life stages have been shown to secrete a number of highly glycosylated molecules [1] and [2]. The SAP of Leishmania mexicana, a component found in large amounts in the culture supernatant of promastigotes, is the best studied example of this group of proteins. It is a copolymer of 100 kDa (SAP1) and 200 kDa (SAP2) subunits which assemble into long filaments [3]. The backbone of the filaments is formed by a ‘string-of-pearls-like’ arrangement of globular protein subunits [4] and [5]. Projecting from this filament, two types of side arms differing in length have been demonstrated by glycerol spraying/low-angle rotary metal shadowing conferring an overall ‘bottle-brush-like’ appearance [5]. These side arms correspond to the Ser/Thr-rich repeat regions of 32 and 383 amino acids of SAP1 and SAP2, respectively, which have been shown to be the exclusive targets for glycosylation via phosphoserines, and a conserved COOH-terminal domain of 54 amino acids [3] and [6]. The composition of the phosphoserine-linked glycans has been studied in detail using monoclonal antibodies directed against phosphodi- and phosphotrisaccharide subunits and oligomannose cap structures [7] and chemical structure analysis [6]. These and other studies showed that SAP shares carbohydrate epitopes with LPG, the major glycolipid component of the promastigote surface [6], [7], [8] and [9]. Here, we show that the Ser/Thr-rich repetitive region of the secreted acid phosphatase of L. mexicana serves as a target for species-specific phosphoglycosylation in Leishmania major.

AB - Parasitic protozoa of the genus Leishmania cycle between two main developmental stages, the promastigote in the digestive tract of the sandfly and the amastigote in the mononuclear cells of the mammalian host. Both life stages have been shown to secrete a number of highly glycosylated molecules [1] and [2]. The SAP of Leishmania mexicana, a component found in large amounts in the culture supernatant of promastigotes, is the best studied example of this group of proteins. It is a copolymer of 100 kDa (SAP1) and 200 kDa (SAP2) subunits which assemble into long filaments [3]. The backbone of the filaments is formed by a ‘string-of-pearls-like’ arrangement of globular protein subunits [4] and [5]. Projecting from this filament, two types of side arms differing in length have been demonstrated by glycerol spraying/low-angle rotary metal shadowing conferring an overall ‘bottle-brush-like’ appearance [5]. These side arms correspond to the Ser/Thr-rich repeat regions of 32 and 383 amino acids of SAP1 and SAP2, respectively, which have been shown to be the exclusive targets for glycosylation via phosphoserines, and a conserved COOH-terminal domain of 54 amino acids [3] and [6]. The composition of the phosphoserine-linked glycans has been studied in detail using monoclonal antibodies directed against phosphodi- and phosphotrisaccharide subunits and oligomannose cap structures [7] and chemical structure analysis [6]. These and other studies showed that SAP shares carbohydrate epitopes with LPG, the major glycolipid component of the promastigote surface [6], [7], [8] and [9]. Here, we show that the Ser/Thr-rich repetitive region of the secreted acid phosphatase of L. mexicana serves as a target for species-specific phosphoglycosylation in Leishmania major.

KW - acid phosphatase

KW - fluorescent antibody technique

KW - glycosylation

KW - immunoblotting

KW - Leishmania major

KW - Leishmania mexicana

KW - plasmids

KW - recombinant proteins

KW - species specificity

KW - secreted acid phosphatase

KW - phosphoglycan modification

KW - filament formation

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DO - 10.1016/S0166-6851(99)00095-X

M3 - Article

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EP - 329

JO - Molecular and Biochemical Parasitology

JF - Molecular and Biochemical Parasitology

SN - 0166-6851

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