To be specific I mean the structure of the human cell
I have never taken a biology class and jumped straight into ap bio this year. I was doing well in chem and jump into ap chem and was told to take this class as well. Admittedly I am confused on a few aspects as reading through some things didn't really help me. Are there any good guides or youtube videos that can explain a bit more on concepts?
Macromolecular drug delivery vectors must translocate at the plasma membrane or escape from endosomes to reach the cytosol. This very inefficient process requires optimization, which is hindered by the difficulty to accurately measure cytosolic arrival. An exceptionally sensitive and robust assay to quantify this step was developed. The assay was validated by determining the absolute numbers of two macromolecular vectors reaching the cytosol.
Abstract
Macromolecular drugs inefficiently cross membranes to reach their cytosolic targets. They require drug delivery vectors to facilitate their translocation across the plasma membrane or escape from endosomes. Optimization of these vectors has however been hindered by the difficulty to accurately measure cytosolic arrival. We have developed an exceptionally sensitive and robust assay for the relative or absolute quantification of this step. The assay is based on benzylguanine and biotin modifications on a drug delivery vector of interest, which allow, respectively, for selective covalent capture in the cytosol with a SNAP-tag fusion protein and for quantification at picomolar sensitivity. The assay was validated by determining the absolute numbers of cytosolic molecules for two drug delivery vectors: the B-subunit of Shiga toxin and the cell-penetrating peptide TAT. We expect this assay to favor delivery vector optimization and the understanding of the enigmatic translocation process.
https://ift.tt/2Ps7LT7
I read that translation happens in the cytoplasm, but also read elsewhere that it happens in the rough ER. Which is it? Does it happen in the cytosol outside of all organelles, by ribosomes floating in the cytosol, or is it in the rough ER by the ribosomes on the ER?
Journal of the American Chemical SocietyDOI: 10.1021/jacs.1c00258
Jessica A. Kretzmann, David C. Luther, Cameron W. Evans, Taewon Jeon, William Jerome, Sanjana Gopalakrishnan, Yi-Wei Lee, Marck Norret, K. Swaminathan Iyer, and Vincent M. Rotello
https://ift.tt/30zdxnR
I'm trying to image a protein that has dual cytosolic and membrane-bound (via protein-protein interaction) localization, but I need to wash out or otherwise not see the cytosolic pool - I'm only interested in seeing the membrane-bound pool. I've tried using digitonin (0.025% in PBS for 10 seconds, then fixation in 4% PFA in PBS for 15 minutes at 37 ℃), but all of the cells (HeLas) ball up and die. I may try a lower digitonin concentration, but I'd like an alternative as well.
Would fixation in ice-cold methanol work for this? I need to be able to see the membrane-associated pool of my protein of interest, as well as a membrane-anchored protein marker.
Any advice on fixation methods for this would be much appreciated!
This may seem silly but one of the things that I cannot ever get straight is confidently stating where a process takes place, or what is located within one of these spaces. I’m specifically talking about the cytosol, cytoplasm, ECS, ICS, etc. I seem to always forget after a couple weeks of getting a card correct.
Does anyone have a good explanation, or ways to remember how to differentiate?
Can someone explain me these cards please? https://imgur.com/a/i0LmTcQ and this https://imgur.com/a/g0roPVv Why differentiate this? TIA
Macromolecular drugs inefficiently cross membranes to reach their cytosolic targets. They require drug delivery vectors to facilitate their translocation across the plasma membrane or escape from endosomes. Optimization of these vectors has however been hindered by the difficulty to accurately measure cytosolic arrival. We have developed an exceptionally sensitive and robust assay for the relative or absolute quantification of this step. The assay is based on benzylguanine and biotin modifications on a drug delivery vector of interest, which allow respectively for selective covalent capture in the cytosol with a SNAP‐tag fusion protein and for quantification at picomolar sensitivity. The assay was validated by determining the absolute numbers of cytosolic molecules for two drug delivery vectors: The B‐subunit of Shiga toxin and the cell‐penetrating peptide TAT. We expect this assay to favour delivery vector optimization and the understanding of the enigmatic translocation process.
https://ift.tt/2Ps7LT7
