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Osteoclasts are cells responsible for bone resorption and remodeling. They originate from hematopoietic progenitor cells present in the bone marrow. Numerous progenitor cells fuse to form multinucleated cells, each with 10-20 nuclei. A single osteoclast has a diameter of 150 to 200 µM. These cells have ruffled borders that break down the underlying bone tissue and release minerals such as calcium into the blood in bone resorption. Osteoclasts cling to bones with their ruffled edges during...
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Bone remodeling is a continuous and balanced process of bone resorption by osteoclasts and bone formation by osteoblasts. In adults, it helps maintain bone mass and calcium homeostasis. While mechanical stress can stimulate turnover as part of the normal maintenance and reparative process, several hormones also regulate bone remodeling.
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Chromatin modification alters gene expression; therefore, scientists can add histone-modifying enzymes, histone variants, and chromatin remodeling complexes to somatic cells to aid reprogramming into pluripotent stem (iPS) cells.
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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
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The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
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Remodeling KRAS.

Daniel J Deredge1, Patrick L Wintrode1

  • 1Department of Pharmaceutical Sciences, University of Maryland School of Pharmacy, 20 North Pine St., Baltimore, MD 21201, USA.

Structure (London, England : 1993)
|September 7, 2017
PubMed
Summary

Targeting cancer-driving RAS proteins is challenging. This study uses hydrogen-deuterium exchange mass spectrometry (HDX-MS) to analyze compounds affecting an oncogenic KRAS mutant, aiding drug development.

Area of Science:

  • Oncogenic signaling pathways
  • Protein structure and dynamics
  • Drug discovery and development

Background:

  • Mutations in RAS family small GTPases are implicated in various human cancers.
  • RAS proteins are historically difficult to target therapeutically due to their structure and function.
  • Oncogenic KRAS mutations drive tumor growth and represent a significant unmet medical need.

Purpose of the Study:

  • To investigate the structural and dynamic effects of two distinct chemical compounds on an oncogenic KRAS mutant.
  • To evaluate the utility of Hydrogen-Deuterium Exchange Mass Spectrometry (HDX-MS) as a tool for characterizing drug interactions with RAS proteins.
  • To provide insights into potential strategies for targeting KRAS-driven cancers.

Main Methods:

  • Utilized Hydrogen-Deuterium Exchange Mass Spectrometry (HDX-MS) to monitor protein structural dynamics.

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  • Analyzed the interaction of two small molecules with different chemical scaffolds against a specific oncogenic KRAS mutant.
  • Characterized changes in protein structure and dynamics upon compound binding.

    • Main Results:

    • Observed distinct effects of the two compounds on the structure and dynamics of the oncogenic KRAS mutant.
    • HDX-MS successfully captured compound-induced conformational changes in KRAS.
    • The study identified specific regions of KRAS affected by the tested compounds.

    Conclusions:

    • HDX-MS is a valuable technique for studying the effects of small molecules on RAS protein structure and dynamics.
    • Understanding these interactions is crucial for developing effective therapies against KRAS-mutated cancers.
    • The findings support the continued development of novel KRAS-targeting drugs.