RESUMEN
A challenge within the field of bioconjugation is developing probes to uncover novel information on proteins and other biomolecules. Intracellular delivery of these probes offers the promise of giving relevant context to this information, and these probes can serve as hypothesis-generating tools within complex systems. Leveraging the utility of triazabutadiene chemistry, herein, we discuss the development of a probe that undergoes reduction-mediated deprotection to rapidly deliver a benzene diazonium ion (BDI) into cells. The intracellular BDI resulted in an increase in global tyrosine phosphorylation levels. Seeing phosphatase dysregulation as a potential source of this increase, a tyrosine phosphatase (PTP1B) was tested and shown to be both inhibited and covalently modified by the BDI. In addition to the expected azobenzene formation at tyrosine side chains, key reactive histidine residues were also modified.
Asunto(s)
Benceno , Proteína Tirosina Fosfatasa no Receptora Tipo 1 , Iones , Fosforilación , Proteína Tirosina Fosfatasa no Receptora Tipo 1/metabolismo , Proteínas/química , Tirosina/metabolismoRESUMEN
IMPORTANCE: Summer Research Experiences for Undergraduates (REUs) are established to provide platforms for interest in scientific research and as tools for eventual matriculation to scientific graduate programs. Unfortunately, the COVID-19 pandemic forced the cancellation of in-person programs for 2020 and 2021, creating the need for alternative programming. The National Summer Undergraduate Research Project (NSURP) was created to provide a virtual option to REUs in microbiology to compensate for the pandemic-initiated loss of research opportunities. Although in-person REUs have since been restored, NSURP currently remains an option for those unable to travel to in-person programs in the first place due to familial, community, and/or monetary obligations. This study examines the effects of the program's first 3 years, documenting the students' experiences, and suggests future directions and areas of study related to the impact of virtual research experiences on expanding and diversifying science, technology, engineering, and mathematics.
Asunto(s)
Disciplinas de las Ciencias Biológicas , Tutoría , Humanos , Pandemias , Mentores , EstudiantesRESUMEN
Nontoxic cryoprotectants are needed for storage of tissues and food preservation. Frozen tissue is particularly susceptible to damage caused by formation of large ice crystals during the thawing process. The current practice of using 5 wt % DMSO for cryopreservation does not produce 100% cell viability post-thaw, at least in part because of DMSO toxicity that is manifested during the freezing and thawing stages of the process. Recently, poly(vinyl alcohol) (PVA) has shown promise in inhibiting ice recrystallization, an activity that is critical for cryoprotection. Inspired by this discovery, we have evaluated nylon-3 polymers for ice recrystallization inhibition activity and for toxicity toward mammalian cells. A survey of homo- and heteropolymers, with side chains bearing variable functionality, has identified new nylon-3 materials that display excellent ice recrystallization inhibition activity and low toxicity.