Pulse marking

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Pulse marking for 30 min. with observation after ten days

The pulse-labeling ( English labeling pulse, pulse-chase analysis ) is a biochemical method for the labeling and tracking of biomolecules . It is a form of metabolic marker .

properties

The pulse marking is used to determine a time course of a molecule in a complex mixture, e.g. B. in an organism, a cell , a lysate or a defined mixture of enzymes . The marking, usually an isotope marking, is carried out by adding the signaling molecule (the pulse ) for a limited time (usually a few minutes ). Then the conditions before the marking ( chase 'follow-up') are restored, e.g. B. by replacing the culture medium used for marking by a culture medium with the same, but unmarked components. For the passage of time, samples are taken at certain time intervals. The metabolism of the labeled molecules or the biosynthesis of molecules generated only after the time of addition can be observed. The tracked molecule can e.g. B. be a metabolite , a protein , a nucleic acid or a carbohydrate .

Applications

The pulse marking enabled various biochemical processes to be further elucidated, e.g. B. translation , protein kinase C , proteasome , and the assembly of bacteriophage T4. With the pulse marking, George Palade was able to elucidate the secretion through the rough endoplasmic reticulum and the Golgi apparatus , for which he received the 1974 Nobel Prize in Physiology or Medicine (“for her discoveries about the structural and functional organization of the cell”).

Individual evidence

  1. ↑ F. GROS, H. Hiatt, W. GILBERT, CG KURLAND, RW Risebrough, JD WATSON: Unstable ribonucleic acid revealed by pulse labeling of Escherichia coli. In: Nature. Volume 190, May 1961, pp. 581-585, ISSN  0028-0836 . PMID 13708983 .
  2. ↑ M. Takahashi, Y. Ono: Pulse-chase analysis of protein kinase C. In: Methods in molecular biology (Clifton, NJ). Volume 233, 2003, pp. 163-170, ISSN  1064-3745 . doi : 10.1385 / 1-59259-397-6: 163 . PMID 12840506 .
  3. ^ MA Hoyt, J. Zich, J. Takeuchi, M. Zhang, C. Govaerts, P. Coffino: Glycine-alanine repeats impair proper substrate unfolding by the proteasome. In: The EMBO journal. Volume 25, Number 8, April 2006, pp. 1720-1729, ISSN  0261-4189 . doi : 10.1038 / sj.emboj.7601058 . PMID 16601692 . PMC 1440830 (free full text).
  4. ↑ PL Ferguson, DH Coombs: Pulse-chase analysis of the in vivo assembly of the bacteriophage T4 tail. In: Journal of molecular biology. Volume 297, Number 1, March 2000, pp. 99-117, ISSN  0022-2836 . doi : 10.1006 / jmbi.2000.3551 . PMID 10704310 .
  5. ^ JD Castle, JD Jamieson, GE Palade: Radioautographic analysis of the secretory process in the parotid acinar cell of the rabbit. In: Journal of Cell Biology . Volume 53, Number 2, May 1972, pp. 290-311, ISSN  0021-9525 . PMID 5025103 . PMC 2108718 (free full text).
  6. ↑ LG CARO, GE PALADE: PROTEIN SYNTHESIS, STORAGE, AND DISCHARGE IN THE PANCREATIC EXOCRINE CELL. TO AUTORADIOGRAPHIC STUDY. In: Journal of Cell Biology . Volume 20, March 1964, pp. 473-495, ISSN  0021-9525 . PMID 14128049 . PMC 2106415 (free full text).