Nucleo-cytoplasmic Transport Assay

In both normal and malignant tissues, specific proteins shuttling from the nucleus are essential for the regulation of the cell cycle and proliferation. Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease affecting the motor neurons and resulting in paralysis and ultimately in death, within 2-5 years on average. Many studies have shown a connection between ALS and impairments in the nucleocytoplasmic pathway. Creative Biolabs is flexible to meet the unique needs of client neurology projects. We can apply our considerable experience with nucleo-cytoplasmic transport assay to develop the necessary analytics specific to your project.

Overview of Nucleo-cytoplasmic Transport (NCT)

NCT refers to the import and export of large molecules from the cell nucleus. Nucleocytoplasmic exchange of proteins and RNAs is mediated by receptors, guiding their cargo through nuclear pores. The peptide locator signal on each cargo determines the receptor with which it will interact. So far, three types of transporters have been identified. The best-studied group is the import protein-β-like protein family (import and export proteins, also known as nuclides). The second class comprises the small homodimeric nuclear transport factor 2 (NTF2)/p10, which imports the small GTPase Ran into the nucleus. The final group is the Tap/Mex67 family, which is involved in mRNA transport.

Fig.1 A model for PABP NCT. (Burgess & Gray, 2012)

General Workflow of NCT at Creative Biolabs

This workflow describes a new and very sensitive assay to evaluate and quantify NCT dysfunction in real-time. The import rate of NLS-NES-GFP protein (shuttle-GFP) can be quantified in real-time by fluorescence microscopy. This is done using an exportin inhibitor, thus allowing the shuttle-GFP only to enter the nucleus.

Fig.2 General workflow of NCT.

Impaired NCT is One Fundamental Pathogenesis of Neurodegenerative Diseases (NDs)

Accumulating evidence indicates that alterations of NCT are the fundamental pathological factors underlying these NDs. Under normal NCT conditions, the protein transport mechanisms enable each protein cargo to reach and retain in appropriate compartments, nucleoplasm, or cytoplasm. In a healthy neuron, most of the mRNAs should be exported to the cytoplasm for protein synthesis. Dysregulations of the protein NCT process will interfere with the normal distributions of protein cargos, leading to protein mislocalization. Protein mislocalization may occur in the nucleus or cytoplasm in diseased or aged neurons. If nuclear transcript export mechanisms are impaired in a neuron, the normal distribution of RNAs will be disrupted, such as nuclear RNA accumulation and mRNA mislocalization. Neurons are specialized cellular subtypes that possess unique features and functions, these features make neurons more vulnerable to the impairment of intracellular transport. The implication of defective NCT in NDs has been revealed by a variety of research model systems.

NCT Assay Services at Creative Biolabs

Nuclear import and export are a highly-coordinated process involving many proteins and large complexes working together on the nuclear membrane. Creative Biolabs provides flexible, robust, facile, and highly amenable to academic scale research assays with the potential to be employed in novel drug-screening applications, in particular, when combined with biochemical assays.

Creative Biolabs is 100% dedicated to the neurology research industry. Our team of highly qualified and experienced technical staff will work with you to develop and deliver assay solutions to add value to your product or project. Please feel free to contact us to discuss your needs.

Reference

1. Burgess, H.M.; Gray, N.K. An integrated model for the nucleo-cytoplasmic transport of cytoplasmic poly (A)-binding proteins. Communicative & integrative biology. 2012, 5(3): 243-247.