Asian journal of andrology, 2011-03, Vol.13 (2), p.266-274
2011
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- Autor(en) / Beteiligte
- Titel
- CABYR binds to AKAP3 and Ropporin in the human sperm fibrous sheath
- Ist Teil von
- Asian journal of andrology, 2011-03, Vol.13 (2), p.266-274
- Ort / Verlag
- China: Medknow Publications & Media Pvt. Ltd
- Erscheinungsjahr
- 2011
- Quelle
- MEDLINE
- Beschreibungen/Notizen
- Calcium-binding tyrosine phosphorylation-regulated protein （CABYR） is a highly polymorphic calcium-binding tyrosine- and serine-/threonine-phosphorylated fibrous sheath （FS） protein involved in capacitation. A putative domain （amino acids 12-48） homologous to the regulatory subunit of type II cAMP-dependent protein kinase A （RII） dimerisation and A kinase-anchoring protein （AKAP）-binding domains of protein kinase A at the N-terminus suggests that CABYR may self-assemble and bind to AKAPs. Moreover, there is evidence that CABYR has limited interaction with AKAPs. However, further evidence and new relationships between CABYR and other FS proteins, including AKAPs, will be helpful in understanding the basic physiology of FS. In this study, a new strategy for co-immunoprecipitation of insoluble proteins, as well as the standard co-immunoprecipitation method in combination with mass spectrometry and western blot, was employed to explore the relationship between CABYR, AKAP3 and Ropperin. The results showed that AKAP3 was co.immunoprecipitated with CABYR by the anti-CABYR-A polyclonal antibody, and, conversely, CABYR was also co.immunoprecipitated with AKAP3 by the anti-AKAP3 polyclonal antibody. Another RIl-like domain containing protein, Ropporin, was also co-immunoprecipitated with CABYR, indicating that Ropporin is one of CABYR＇s binding partners. The interactions between CABYR, AKAP3 and Ropporin were confirmed by yeast two-hybrid assays. Further analysis showed that CABYR not only binds to AKAP3 by its RII domain but binds to Ropporin through other regions besides the RIl-like domain. This is the first demonstration that CABYR variants form a complex not only with the scaffolding protein AKAP3 but also with another Rll-like domain-containing protein in the human sperm FS.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 1008-682XeISSN: 1745-7262DOI: 10.1038/aja.2010.149Titel-ID: cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_3739192
- Format
- –
- Schlagworte
- A Kinase Anchor Proteins - chemistry, A Kinase Anchor Proteins - genetics, A Kinase Anchor Proteins - metabolism, Amino Acid Sequence, Calcium-Binding Proteins - chemistry, Calcium-Binding Proteins - genetics, Calcium-Binding Proteins - metabolism, Genetic Variation, Humans, Immunoprecipitation, Male, Membrane Proteins - chemistry, Membrane Proteins - genetics, Membrane Proteins - metabolism, Molecular Sequence Data, Multiprotein Complexes - chemistry, Original, Phosphoproteins - chemistry, Phosphoproteins - genetics, Phosphoproteins - metabolism, Protein Binding, Protein Interaction Domains and Motifs, Recombinant Fusion Proteins - chemistry, Recombinant Fusion Proteins - genetics, Recombinant Fusion Proteins - metabolism, rho GTP-Binding Proteins - chemistry, rho GTP-Binding Proteins - genetics, rho GTP-Binding Proteins - metabolism, Sperm Tail - metabolism, Sperm Tail - ultrastructure, Two-Hybrid System Techniques, 免疫共沉淀, 多克隆抗体, 蛋白激酶A, 调节蛋白, 酪氨酸磷酸化