scholarly journals Genetically encoded live-cell sensor for tyrosinated microtubules

2020 ◽  
Vol 219 (10) ◽  
Author(s):  
Shubham Kesarwani ◽  
Prakash Lama ◽  
Anchal Chandra ◽  
P. Purushotam Reddy ◽  
A.S. Jijumon ◽  
...  
Keyword(s):  
Real Time ◽  
Posttranslational Modifications ◽  
Living Cells ◽  
Live Cell ◽  
Yeast Display ◽  
Cellular Functions ◽  
Microtubule Stability ◽  
Specific Manner ◽  
Potential Applications ◽  
Stability Dynamics

Microtubule cytoskeleton exists in various biochemical forms in different cells due to tubulin posttranslational modifications (PTMs). Tubulin PTMs are known to affect microtubule stability, dynamics, and interaction with MAPs and motors in a specific manner, widely known as tubulin code hypothesis. At present, there exists no tool that can specifically mark tubulin PTMs in living cells, thus severely limiting our understanding of their dynamics and cellular functions. Using a yeast display library, we identified a binder against terminal tyrosine of α-tubulin, a unique PTM site. Extensive characterization validates the robustness and nonperturbing nature of our binder as tyrosination sensor, a live-cell tubulin nanobody specific towards tyrosinated microtubules. Using this sensor, we followed nocodazole-, colchicine-, and vincristine-induced depolymerization events of tyrosinated microtubules in real time and found each distinctly perturbs the microtubule polymer. Together, our work describes a novel tyrosination sensor and its potential applications to study the dynamics of microtubule and their PTM processes in living cells.

scholarly journals Genetically encoded live cell sensor for tyrosinated microtubules

2020 ◽  
Author(s):  
Shubham Kesarwani ◽  
Prakash Lama ◽  
Anchal Chandra ◽  
P. Purushotam Reddy ◽  
AS Jijumon ◽  
...  
Keyword(s):  
Living Cells ◽  
Live Cell ◽  
Live Cells ◽  
Yeast Display ◽  
Post Translational Modification ◽  
Alpha Tubulin ◽  
Microtubule Stability ◽  
Specific Manner ◽  
Potential Applications ◽  
Stability Dynamics

AbstractMicrotubule cytoskeleton exists in various biochemical forms in different cells due to tubulin post-translational modification (PTMs). These PTMs are known to affect microtubule stability, dynamics and interaction with MAPs and motors in a specific manner, widely known as tubulin code hypothesis. At present there exist no tool that can specifically mark tubulin PTMs in live cells, thus severely limiting our understanding of tubulin PTMs. Using yeast display library, we identified a binder against terminal tyrosine of alpha tubulin, a unique PTM site. Extensive characterization validates the robustness and non-perturbing nature of our binder as tyrosination sensor, a live cell tubulin nanobody specific towards tyrosinated or unmodified microtubules. Using which, in real time we followed nocodazole, colchicine and vincristine induced depolymerization events of unmodified microtubules, and found each distinctly perturb microtubule polymer. Together, our work describes the tyrosination sensor and potential applications to study microtubule and PTM processes in living cells.

BODIPY based realtime, reversible and targeted fluorescent probes for biothiol imaging in living cells

2020 ◽  
Vol 56 (93) ◽  
pp. 14717-14720
Author(s):  
Rongkun He ◽  
Yichuan Zhang ◽  
Suresh Madhu ◽  
Quan Gao ◽  
Qianjin Lian ◽  
...  
Keyword(s):  
Real Time ◽  
Fluorescent Probes ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Living Cells ◽  
Live Cell

Real-time live cell imaging and quantification of biothiol dynamics are important for understanding pathophysiological processes.

Multimode light microscopy and fluorescence-based reagents as tools for the study of chemical and molecular dynamics of living cells and tissues

1992 ◽  
Vol 50 (1) ◽  
pp. 418-419
Author(s):  
D. L. Taylor
Keyword(s):  
Living Cells ◽  
Cellular Level ◽  
Molecular Techniques ◽  
Cellular Functions ◽  
Macromolecular Assemblies ◽  
Cell Functions ◽  
Molecular Events ◽  
Yield Information ◽  
Full Knowledge ◽  
Structural Methods

Cells function through the complex temporal and spatial interplay of ions, metabolites, macromolecules and macromolecular assemblies. Biochemical approaches allow the investigator to define the components and the solution chemical reactions that might be involved in cellular functions. Static structural methods can yield information concerning the 2- and 3-D organization of known and unknown cellular constituents. Genetic and molecular techniques are powerful approaches that can alter specific functions through the manipulation of gene products and thus identify necessary components and sequences of molecular events. However, full knowledge of the mechanism of particular cell functions will require direct measurement of the interplay of cellular constituents. Therefore, there has been a need to develop methods that can yield chemical and molecular information in time and space in living cells, while allowing the integration of information from biochemical, molecular and genetic approaches at the cellular level.

Recent Developments of Ambulatory Assessment Methods

2014 ◽  
Vol 25 (4) ◽  
pp. 279-287 ◽  
Author(s):  
Stefan Hey ◽  
Panagiota Anastasopoulou ◽  
André Bideaux ◽  
Wilhelm Stork
Keyword(s):  
Real Time ◽  
Information And Communication Technologies ◽  
New Technologies ◽  
Psychological Research ◽  
Ambulatory Assessment ◽  
Emotional States ◽  
Recent Developments ◽  
Potential Applications ◽  
Information And Communication ◽  
Physiological Sensors

Ambulatory assessment of emotional states as well as psychophysiological, cognitive and behavioral reactions constitutes an approach, which is increasingly being used in psychological research. Due to new developments in the field of information and communication technologies and an improved application of mobile physiological sensors, various new systems have been introduced. Methods of experience sampling allow to assess dynamic changes of subjective evaluations in real time and new sensor technologies permit a measurement of physiological responses. In addition, new technologies facilitate the interactive assessment of subjective, physiological, and behavioral data in real-time. Here, we describe these recent developments from the perspective of engineering science and discuss potential applications in the field of neuropsychology.

PUF Proteins: Cellular Functions and Potential Applications

2017 ◽  
Vol 18 (3) ◽  
pp. 250-261 ◽  
Author(s):  
Seyed Kiani ◽  
Tahereh Taheri ◽  
Sima Rafati ◽  
Katayoun Samimi-Rad
Keyword(s):  
Cellular Functions ◽  
Puf Proteins ◽  
Potential Applications

scholarly journals Tubulin glycylation controls axonemal dynein activity, flagellar beat, and male fertility

Science ◽  
2021 ◽  
Vol 371 (6525) ◽  
pp. eabd4914
Author(s):  
Sudarshan Gadadhar ◽  
Gonzalo Alvarez Viar ◽  
Jan Niklas Hansen ◽  
An Gong ◽  
Aleksandr Kostarev ◽  
...  
Keyword(s):  
Posttranslational Modifications ◽  
Male Fertility ◽  
Electron Tomography ◽  
Structural Basis ◽  
Cellular Functions ◽  
Flagellar Beat ◽  
Cryo Electron Tomography ◽  
Axonemal Dynein ◽  
Dynein Arms ◽  
Cilia And Flagella

Posttranslational modifications of the microtubule cytoskeleton have emerged as key regulators of cellular functions, and their perturbations have been linked to a growing number of human pathologies. Tubulin glycylation modifies microtubules specifically in cilia and flagella, but its functional and mechanistic roles remain unclear. In this study, we generated a mouse model entirely lacking tubulin glycylation. Male mice were subfertile owing to aberrant beat patterns of their sperm flagella, which impeded the straight swimming of sperm cells. Using cryo–electron tomography, we showed that lack of glycylation caused abnormal conformations of the dynein arms within sperm axonemes, providing the structural basis for the observed dysfunction. Our findings reveal the importance of microtubule glycylation for controlled flagellar beating, directional sperm swimming, and male fertility.

Sign in / Sign up

Export Citation Format

Share Document