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A nanobody suite for yeast scaffold nucleoporins provides details of the nuclear pore complex structure
- Source :
- Nature Communications, Nature Communications, Vol 11, Iss 1, Pp 1-13 (2020), Nordeen, S A, Andersen, K R, Knockenhauer, K E, Ingram, J R, Ploegh, H L & Schwartz, T U 2020, ' A nanobody suite for yeast scaffold nucleoporins provides details of the nuclear pore complex structure ', Nature Communications, vol. 11, no. 1, 6179 . https://doi.org/10.1038/s41467-020-19884-6
- Publication Year :
- 2020
-
Abstract
- Nuclear pore complexes (NPCs) are the main conduits for molecular exchange across the nuclear envelope. The NPC is a modular assembly of ~500 individual proteins, called nucleoporins or nups. Most scaffolding nups are organized in two multimeric subcomplexes, the Nup84 or Y complex and the Nic96 or inner ring complex. Working in S. cerevisiae, and to study the assembly of these two essential subcomplexes, we here develop a set of twelve nanobodies that recognize seven constituent nucleoporins of the Y and Nic96 complexes. These nanobodies all bind specifically and with high affinity. We present structures of several nup-nanobody complexes, revealing their binding sites. Additionally, constitutive expression of the nanobody suite in S. cerevisiae detect accessible and obstructed surfaces of the Y complex and Nic96 within the NPC. Overall, this suite of nanobodies provides a unique and versatile toolkit for the study of the NPC.<br />Characterizing the assembly of the nuclear pore complex (NPC) remains challenging. Here, the authors develop a set of nanobodies that recognize seven constituent nucleoporins, study their binding characteristics, and apply them to probe accessible and obstructed NPC surfaces in yeast.
- Subjects :
- 0301 basic medicine
Models, Molecular
Protein Conformation, alpha-Helical
Scaffold
Beta sheet
Antibody Affinity
General Physics and Astronomy
Gene Expression
Plasma protein binding
Crystallography, X-Ray
0302 clinical medicine
Protein structure
Antibody Specificity
Protein Isoforms
Nuclear pore
Cloning, Molecular
Multidisciplinary
Chemistry
Recombinant Proteins
Nucleoporin
Camelids, New World
Molecular exchange
Protein Binding
Saccharomyces cerevisiae Proteins
Science
Genetic Vectors
Sequence alignment
Saccharomyces cerevisiae
General Biochemistry, Genetics and Molecular Biology
Article
03 medical and health sciences
Peptide Library
otorhinolaryngologic diseases
Escherichia coli
Animals
Protein Interaction Domains and Motifs
Amino Acid Sequence
Binding site
Peptide library
X-ray crystallography
Binding Sites
Sequence Homology, Amino Acid
Proteins
General Chemistry
Single-Domain Antibodies
Yeast
Nuclear Pore Complex Proteins
Nuclear pore complex
stomatognathic diseases
Kinetics
030104 developmental biology
Biophysics
Nuclear Pore
Protein Conformation, beta-Strand
Protein design
Sequence Alignment
030217 neurology & neurosurgery
Subjects
Details
- ISSN :
- 20411723
- Volume :
- 11
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- Nature communications
- Accession number :
- edsair.doi.dedup.....e41864a8cf9289358431daaade8ccb4e