Recyclable chaperone-conjugated magnetic beads for in vitro refolding of Burkholderia cepacia lipase

Poly(ADP-ribose) (pADPr), made by PARP-5a/tankyrase-1, localizes to the poles of mitotic spindles and is required for bipolar spindle assembly, but its molecular function in the spindle is poorly understood. To investigate this, we localized pADPr at spindle poles by immuno-EM. We then developed a concentrated mitotic lysate system from HeLa cells to probe spindle pole assembly in vitro. Microtubule asters assembled in response to centrosomes and Ran-GTP in this system. Magnetic beads coated with pADPr, extended from PARP-5a, also triggered aster assembly, suggesting a functional role of the pADPr in spindle pole assembly. We found that PARP-5a is much more active in mitosis than interphase. We used mitotic PARP-5a, self-modified with pADPr chains, to capture mitosis-specific pADPr-binding proteins. Candidate binding proteins included the spindle pole protein NuMA previously shown to bind to PARP-5a directly. The rod domain of NuMA, expressed in bacteria, bound directly to pADPr. We propose that pADPr provides a dynamic cross-linking function at spindle poles by extending from covalent modification sites on PARP-5a and NuMA and binding noncovalently to NuMA and that this function helps promote assembly of exactly two poles.

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Molecular mechanism of activation of Burkholderia cepacia lipase at aqueous–organic interfaces

Physical Chemistry Chemical Physics ◽

10.1039/c7cp04466f ◽

2017 ◽

Vol 19 (46) ◽

pp. 31499-31507 ◽

Cited By ~ 5

Author(s):

Ivan Pires de Oliveira ◽

Gabriel Ernesto Jara ◽

Leandro Martínez

Keyword(s):

Molecular Mechanism ◽

Burkholderia Cepacia ◽

Burkholderia Cepacia Lipase ◽

Organic Interfaces

Structure and thermodynamics of lipase activation at aqueous–organic interfaces.

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In Vitro Stimulation and Expansion of Human Tumour-Reactive CD8+ Cytotoxic T Lymphocytes by Anti-CD3/CD28/CD137 Magnetic Beads

Scandinavian Journal of Immunology ◽

10.1111/j.1365-3083.2011.02564.x ◽

2011 ◽

Vol 74 (2) ◽

pp. 155-164 ◽

Cited By ~ 13

Author(s):

D. Teschner ◽

G. Wenzel ◽

E. Distler ◽

E. Schnürer ◽

M. Theobald ◽

...

Keyword(s):

T Lymphocytes ◽

Cytotoxic T Lymphocytes ◽

Magnetic Beads ◽

Human Tumour

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Aptamer-targeting of Aleutian mink disease virus (AMDV) can be an effective strategy to inhibit virus replication

Scientific Reports ◽

10.1038/s41598-021-84223-8 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Taofeng Lu ◽

Hui Zhang ◽

Jie Zhou ◽

Qin Ma ◽

Wenzhuo Yan ◽

...

Keyword(s):

Disease Virus ◽

Magnetic Beads ◽

Inhibitory Effect ◽

Three Dimensions ◽

Economic Losses ◽

Effective Vaccine ◽

Stem Loop ◽

Aleutian Mink Disease Virus ◽

Random Library

AbstractAleutian mink disease (AMD), which is caused by Aleutian mink disease virus (AMDV), is an important contagious disease for which no effective vaccine is yet available. AMD causes major economic losses for mink farmers globally and threatens some carnivores such as skunks, genets, foxes and raccoons. Aptamers have exciting potential for the diagnosis and/or treatment of infectious viral diseases, including AMD. Using a magnetic beads-based systemic evolution of ligands by exponential enrichment (SELEX) approach, we have developed aptamers with activity against AMDV after 10 rounds of selection. After incubation with the ADVa012 aptamer (4 μM) for 48 h, the concentration of AMDV in the supernatant of infected cells was 47% lower than in the supernatant of untreated cells, whereas a random library of aptamers has no effect. The half-life of ADVa012 was ~ 32 h, which is significantly longer than that of other aptamers. Sequences and three dimensions structural modeling of selected aptamers indicated that they fold into similar stem-loop structures, which may be a preferred structure for binding to the target protein. The ADVa012 aptamer was shown to have an effective and long-lasting inhibitory effect on viral production in vitro.

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