Characterization of the interaction of the chemotherapeutic drug mitomycin C with DNA in vitro and in vivo and effects on specific DNA-protein interactions.

1996 ◽  
Author(s):  
Amy J. Warren
Keyword(s):  
Mitomycin C ◽  
Protein Interactions ◽  
Chemotherapeutic Drug ◽  
Dna Protein Interactions

scholarly journals In vivo analysis of DNA-protein interactions on the human erythropoietin enhancer.

1997 ◽  
Vol 17 (2) ◽  
pp. 851-856 ◽  
Author(s):  
B Hu ◽  
E Wright ◽  
L Campbell ◽  
K L Blanchard
Keyword(s):  
Oxygen Tension ◽  
Protein Interactions ◽  
Proximal Promoter ◽  
Low Oxygen ◽  
Initiation Rate ◽  
Cobalt Exposure ◽  
In Cells ◽  
Dna Protein Interactions

The erythropoietin (EPO) gene is one of the best examples of a mammalian gene controlled by oxygen tension. The DNA elements responsible for hypoxia-induced transcription consist of a short region of the proximal promoter and a <50-bp 3' enhancer. The elements act cooperatively to increase the transcriptional initiation rate approximately 100-fold in response to low oxygen tension in Hep3B cells. Two distinct types of transactivating proteins have been demonstrated to bind the response elements in the human EPO enhancer in vitro: one shows hypoxia-inducible DNA binding activity, while the other activity binds DNA under normoxic and hypoxic conditions. We have investigated the DNA-protein interactions on the human EPO enhancer in living tissue culture cells that produce EPO in a regulated fashion (Hep3B) and in cells that do not express EPO under any conditions tested (HeLa). We have identified in vivo DNA-protein interactions on the control elements in the human EPO enhancer by ligation-mediated PCR technology. We show that the putative protein binding sites in the EPO enhancer are occupied in vivo under conditions of normoxia, hypoxia, and cobalt exposure in EPO-producing cells. These sites are not occupied in cells that do not produce EPO. We also provide evidence for a conformational change in the topography of the EPO enhancer in response to hypoxia and cobalt exposure.

scholarly journals The Toxoplasma gondii Cyst Wall Interactome

mBio ◽  
2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Vincent Tu ◽  
Tadakimi Tomita ◽  
Tatsuki Sugi ◽  
Joshua Mayoral ◽  
Bing Han ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Protein Interactions ◽  
Cyst Wall ◽  
Affinity Purification ◽  
Dense Granule ◽  
Hypothetical Proteins ◽  
Content Type

ABSTRACT A characteristic of the latent cyst stage of Toxoplasma gondii is a thick cyst wall that forms underneath the membrane of the bradyzoite vacuole. Previously, our laboratory group published a proteomic analysis of purified in vitro cyst wall fragments that identified an inventory of cyst wall components. To further refine our understanding of the composition of the cyst wall, several cyst wall proteins were tagged with a promiscuous biotin ligase (BirA*), and their interacting partners were screened by streptavidin affinity purification. Within the cyst wall pulldowns, previously described cyst wall proteins, dense granule proteins, and uncharacterized hypothetical proteins were identified. Several of the newly identified hypothetical proteins were validated to be novel components of the cyst wall and tagged with BirA* to expand the model of the cyst wall interactome. Community detection of the cyst wall interactome model revealed three distinct clusters: a dense granule, a cyst matrix, and a cyst wall cluster. Characterization of several of the identified cyst wall proteins using genetic strategies revealed that MCP3 affects in vivo cyst sizes. This study provides a model of the potential protein interactions within the cyst wall and the groundwork to understand cyst wall formation. IMPORTANCE A model of the cyst wall interactome was constructed using proteins identified through BioID. The proteins within this cyst wall interactome model encompass several proteins identified in a prior characterization of the cyst wall proteome. This model provides a more comprehensive understanding of the composition of the cyst wall and may lead to insights on how the cyst wall is formed.

scholarly journals Structural and functional characterization of Rpn12 identifies residues required for Rpn10 proteasome incorporation

2012 ◽  
Vol 448 (1) ◽  
pp. 55-65 ◽  
Author(s):  
Jonas Boehringer ◽  
Christiane Riedinger ◽  
Konstantinos Paraskevopoulos ◽  
Eachan O. D. Johnson ◽  
Edward D. Lowe ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Functional Characterization ◽  
Ubiquitin Proteasome System ◽  
26S Proteasome ◽  
Structural Motif ◽  
Protein Protein Interactions ◽  
Ubiquitin Proteasome

The ubiquitin–proteasome system targets selected proteins for degradation by the 26S proteasome. Rpn12 is an essential component of the 19S regulatory particle and plays a role in recruiting the extrinsic ubiquitin receptor Rpn10. In the present paper we report the crystal structure of Rpn12, a proteasomal PCI-domain-containing protein. The structure helps to define a core structural motif for the PCI domain and identifies potential sites through which Rpn12 might form protein–protein interactions. We demonstrate that mutating residues at one of these sites impairs Rpn12 binding to Rpn10 in vitro and reduces Rpn10 incorporation into proteasomes in vivo.

Biomimetic Nanohybrids for Combined imaging and Cancer Therapy

2012 ◽  
Vol 1468 ◽  
Author(s):  
A. Topete ◽  
P. Iglesias ◽  
M. Alatorre-Meda ◽  
A. Cambón ◽  
E. Alvarez ◽  
...  
Keyword(s):  
Glycolic Acid ◽  
Chemotherapeutic Drug ◽  
Nir Light ◽  
Nir Absorption ◽  
Combined Imaging ◽  
Plga Nanoparticle ◽  
Nir Dye

ABSTRACTIn this work, we have developed a multifunctional theranostic nanoplatform consisting of a poly(lactic-co-glycolic acid) (PLGA) biodegradable matrix covered by a gold shell, which provides the system with NIR absorption ability and subsequent generation of hyperthermia effect. Inside the PLGA nanoparticle, the chemotherapeutic drug doxorubicin (DOXO) and the NIR dye isocyanine green (ICG) were loaded. The characterization of the particles, their in vitro cytotocixity combining NIR light irradiation and chemotherapy and their preliminary in vivo biodistribution is analyzed.

scholarly journals In vivo and in vitro characterization of protein interactions with the dyad G-box of the Arabidopsis Adh gene.

1990 ◽  
Vol 2 (3) ◽  
pp. 207-214 ◽  
Author(s):  
W L McKendree ◽  
A L Paul ◽  
A J DeLisle ◽  
R J Ferl
Keyword(s):  
Protein Interactions ◽  
In Vitro Characterization ◽  
Adh Gene

In vivo and in vitro Characterization of Protein Interactions with the Dyad G-Box of the Arabidopsis Adh Gene

1990 ◽  
Vol 2 (3) ◽  
pp. 207
Author(s):  
William L. McKendree ◽  
Anna-Lisa Paul ◽  
Alice J. DeLisle ◽  
Robert J. Ferl
Keyword(s):  
Protein Interactions ◽  
In Vitro Characterization ◽  
Adh Gene

scholarly journals Identification of in vivo CFTR conformations during biogenesis and upon misfolding by covalent protein painting (CPP)

2021 ◽  
Author(s):  
Sandra Pankow ◽  
Casimir Bamberger ◽  
Salvador Martínez-Bartolomé ◽  
Sung-Kyu Park ◽  
John R. Yates
Keyword(s):  
Cystic Fibrosis ◽  
Protein Interactions ◽  
Structural Changes ◽  
Solvent Accessibility ◽  
Protein Structures ◽  
Permissive Temperature ◽  
Intracellular Loop

AbstractIn vivo characterization of protein structures or protein structural changes after perturbation is a major challenge. Therefore, experiments to characterize protein structures are typically performed in vitro and with highly purified proteins or protein complexes. Using a novel low-resolution method named Covalent Protein Painting (CPP) that allows the characterization of protein conformations in vivo, we are the first to report how an ion channel, the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR), is conformationally changed during biogenesis and channel opening in the cell. Our study led to the identification of a novel opening mechanism for CFTR by revealing that the interaction of the intracellular loop 2 (ICL2) with the nucleotide binding domain 2 (NDB2) of CFTR is needed for channel gating, and this interaction occurs concomitantly with changes to the narrow part of the pore and the walker A lysine in NBD1. However, the ICL2:NBD2 interface, which forms a “ball-in-a-socket” motif, is uncoupled during biogenesis, likely to prevent inadvertent channel activation during transport. In particular, solvent accessibility of lysine 273 (K273) in ICL2 changes with the opening and closing of the channel. Mutation of K273 severely impaired CFTR biogenesis and led to accumulation of CFTR in the Golgi and TGN. CPP further revealed that, even upon treatment with current approved drugs or at permissive temperature, the uncoupled state of ICL2 is a prominent feature of the misfolded CFTR mutants ΔF508 and N1303K that cause Cystic Fibrosis (CF), which suggests that stabilization of this interface could produce a more efficient CF drug. CPP is able to characterize a protein in its native environment and measure the effect of complex PTMs and protein interactions on protein structure, making it broadly applicable and invaluable for the development of new therapies.

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