Korean J Parasitol.  2016 Aug;54(4):461-469. 10.3347/kjp.2016.54.4.461.

Identification of a Novel Microtubule-Binding Protein in Giardia lamblia

• Kim J
• Park SJ

• KMID: 2353576
• DOI: http://doi.org/10.3347/kjp.2016.54.4.461

Abstract

Giardia lamblia is a protozoan that causes diarrheal diseases in humans. Cytoskeletal structures of Giardia trophozoites must be finely reorganized during cell division. To identify Giardia proteins which interact with microtubules (MTs), Giardia lysates were incubated with in vitro-polymerized MTs and then precipitated by ultracentifugation. A hypothetical protein (GL50803_8405) was identified in the precipitated fraction with polymerized MTs and was named GlMBP1 (G. lamblia microtubule-binding protein 1). Interaction of GlMBP1 with MTs was confirmed by MT binding assays using recombinant GlMBP1 (rGlMBP1). In vivo expression of GlMBP1 was shown by a real-time PCR and western blot analysis using anti-rGlMBP1 antibodies. Transgenic G. lamblia trophozoites were constructed by integrating a chimeric gene encoding hemagglutinin (HA)-tagged GlMBP1 into a Giardia chromosome. Immunofluorescence assays of this transgenic G. lamblia, using anti-HA antibodies, revealed that GlMBP1 mainly localized at the basal bodies, axonemes, and median bodies of G. lamblia trophozoites. This result indicates that GlMBP1 is a component of the G. lamblia cytoskeleton.

Keyword

Giardia lamblia; microtubule-binding protein 1

MeSH Terms

Antibodies
Axoneme
Basal Bodies
Blotting, Western
Cell Division
Cytoskeleton
Fluorescent Antibody Technique
Giardia lamblia*
Giardia*
Hemagglutinins
Humans
Microtubules
Polymers
Real-Time Polymerase Chain Reaction
Trophozoites
Antibodies
Hemagglutinins
Polymers