scholarly journals In vitro characterization of the full-length human dynein-1 cargo adaptor BicD2

2022 ◽  
Author(s):  
Robert Fagiewicz ◽  
Corinne Crucifix ◽  
Celia Deville ◽  
Bruno Kieffer ◽  
Yves Nomine ◽  
...  
Keyword(s):  
Conformational Changes ◽  
Protein Complexes ◽  
Cytoplasmic Dynein ◽  
Full Length ◽  
Recombinant Production ◽  
In Vitro Characterization ◽  
Microtubule Motor ◽  
Glial Progenitor Cells ◽  
Development In Vitro

The cargo adaptors are crucial in coupling motor proteins with their respective cargos and regulatory proteins. BicD2 is one of the most prominent examples within the cargo adaptor family. BicD2 is able to recruit the microtubule motor dynein to RNA, viral particles and nuclei. The BicD2-mediated interaction between the nucleus and dynein is implicated in mitosis as well as interkinetic nuclear migration (INM) in radial glial progenitor cells, and neuron precursor migration during embryonic neocortex development. In vitro studies involving full-length cargo adaptors are difficult to perform due to the hydrophobic character, low-expression levels, and intrinsic flexibility of cargo adaptors. Here we report the recombinant production of full-length human BicD2 and confirm its biochemical activity by interaction studies with RanBP2 and cytoplasmic dynein-1. We also describe pH-dependent conformational changes of BicD2 using cryoEM, template-free structure predictions, and biophysical tools. Our results will help defining the biochemical parameters for the invitro reconstitution of higher order BicD2 protein complexes.

Real time imaging reveals a peroxisomal reticulum in living cells

2000 ◽  
Vol 113 (20) ◽  
pp. 3663-3671 ◽  
Author(s):  
M. Schrader ◽  
S.J. King ◽  
T.A. Stroh ◽  
T.A. Schroer
Keyword(s):  
Real Time ◽  
Cytoplasmic Dynein ◽  
Living Cells ◽  
Real Time Imaging ◽  
Peroxisomal Targeting ◽  
Microtubule Motor ◽  
Targeting Signal ◽  
Peroxisomal Targeting Signal

We have directly imaged the dynamic behavior of a variety of morphologically different peroxisomal structures in HepG2 and COS-7 cells transfected with a construct encoding GFP bearing the C-terminal peroxisomal targeting signal 1. Real time imaging revealed that moving peroxisomes interacted with each other and were engaged in transient contacts, and at higher magnification, tubular peroxisomes appeared to form a peroxisomal reticulum. Local remodeling of these structures could be observed involving the formation and detachment of tubular processes that interconnected adjacent organelles. Inhibition of cytoplasmic dynein based motility by overexpression of the dynactin subunit, dynamitin (p50), inhibited the movement of peroxisomes in vivo and interfered with the reestablishment of a uniform distribution of peroxisomes after recovery from nocodazole treatment. Isolated peroxisomes moved in vitro along microtubules in the presence of a microtubule motor fraction. Our data reveal that peroxisomal behavior in vivo is significantly more dynamic and interactive than previously thought and suggest a role for the dynein/dynactin motor in peroxisome motility.

Corrigendum to: Transethosomes of Econazole Nitrate for Transdermal Delivery: Development, In-vitro Characterization, and Ex-vivo Assessment

2021 ◽  
Vol 9 (3) ◽  
pp. 245-245
Author(s):  
Shivani Verma ◽  
Puneet Utreja
Keyword(s):  
Transdermal Delivery ◽  
Ex Vivo ◽  
In Vitro Characterization ◽  
Research Scholar ◽  
Development In Vitro ◽  
Econazole Nitrate

The authors wish to add words “Research Scholar” and “Research Supervisor” to their affiliations [1]. The original article can be found online at https://doi.org/10.2174/2211738506666180813122102

Biodegradable polymeric system for cisplatin delivery: Development, in vitro characterization and investigation of toxicity profile

2014 ◽  
Vol 38 ◽  
pp. 85-93 ◽  
Author(s):  
Noor Alam ◽  
Vaibhav Khare ◽  
Ravindra Dubey ◽  
Ankit Saneja ◽  
Manoj Kushwaha ◽  
...  
Keyword(s):  
Toxicity Profile ◽  
Polymeric System ◽  
In Vitro Characterization ◽  
Development In Vitro

Development, in-vitro characterization and assessment of cosmetic potential of Beta vulgaris extract emulsion

2020 ◽  
Vol 23 ◽  
pp. 100372
Author(s):  
Sidra Huma ◽  
Hajji Muhammad Shoaib Khan ◽  
Muhammad Sohail ◽  
Naveed Akhtar ◽  
Fatima Rasool ◽  
...  
Keyword(s):  
Beta Vulgaris ◽  
In Vitro Characterization ◽  
Development In Vitro

scholarly journals Autokinase Activity of Casein Kinase 1 δ/ε Governs the Period of Mammalian Circadian Rhythms

2019 ◽  
Vol 34 (5) ◽  
pp. 482-496 ◽  
Author(s):  
Gaili Guo ◽  
Kankan Wang ◽  
Shan-Shan Hu ◽  
Tian Tian ◽  
Peng Liu ◽  
...  
Keyword(s):  
Circadian Rhythms ◽  
Conformational Changes ◽  
Catalytic Domain ◽  
Biochemical Properties ◽  
Period Change ◽  
Casein Kinase ◽  
Full Length ◽  
Casein Kinase 1 ◽  
Autokinase Activity

Circadian rhythms exist in nearly all organisms. In mammals, transcriptional and translational feedback loops (TTFLs) are believed to underlie the mechanism of the circadian clock. Casein kinase 1δ/ε (CK1δ/ε) are key kinases that phosphorylate clock components such as PER proteins, determining the pace of the clock. Most previous studies of the biochemical properties of the key kinases CK1ε and CK1δ in vitro have focused on the properties of the catalytic domains from which the autoinhibitory C-terminus has been deleted (ΔC); those studies ignored the significance of self-inhibition by autophosphorylation. By comparing the properties of the catalytic domain of CK1δ/ε with the full-length kinase that can undergo autoinhibition, we found that recombinant full-length CK1 showed a sequential autophosphorylation process that induces conformational changes to affect the overall kinase activity. Furthermore, a direct relationship between the period change and the autokinase activity among CK1δ, CK1ε, and CK1ε-R178C was observed. These data implicate the autophosphorylation activity of CK1δ and CK1ε kinases in setting the pace of mammalian circadian rhythms and indicate that the circadian period can be modulated by tuning the autophosphorylation rates of CK1δ/ε.

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