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Collagens are the most abundant family of extra cellular matrix (ECM) proteins, accounting for two-thirds of the dry mass of adult articular cartilage. Several subtypes have been identified including types II, IX, X, XI, VI, XII, and XIV collagen, however 80 to 90% of the collagen in the body consists of types I, II, and III. Collagen is a long thin protein which consists of three coiled subunits: two α1(I) chains and one α2(I). Type 1 collagen forms part of the major structure of the bone, and mutations in the α1(I) or α2(I) genes lead to-osteogenesis imperfecta, or brittle-bone disease.Collagen is cleaved by mammalian tolloid (mTld) and its splice variant- bone morphogenetic protein 1 (BMP1), a metalloprotease and member of the peptidase M12A family enzymes.This peptide contains a FRET (fluorescence or Fö-rster resonance energy transfer) pair. N-terminal 5-Carboxyfluorescein (5-FAM), a widely used green fluorescent tag which excites at 495 nm and emits at 517 nm, and a 5-Carboxytetramethylrhodamine-(5-TAMRA), present on the side chain of a C-terminal lysine which has been added to the peptide. 5-TAMRA is a widely used fluorescent dye which excites at 546 nm and emits at 579 nm. In the intact peptide TAMRA quenches the fluorescence of 5-FAM, however upon cleavage of the peptide by a protease such as BMP1, the fluorescence of 5-FAM is recovered and can be monitored.

TAMRA标记的说明:

Single isomer 5-Carboxytetramethyl rhodamine (5TAMRA) and mixed-isomer 5/6-TAMRA are fluorophors with excitations at ~543 nm ▉ and emissions of ~572 nm ▉.

TAMRA标记肽的相关文献:

Nanoparticle delivery of immunostimulatory oligonucleotides enhances response to checkpoint inhibitor therapeutics
Buss, Colin G., and Sangeeta N. Bhatia.  Proceedings of the National Academy of Sciences (2020).

USP7 small-molecule inhibitors interfere with ubiquitin binding.
Kategaya, L., Di Lello, P., Rougé, L., Pastor, R., Clark, K.R., Drummond, J., Kleinheinz, T., Lin, E., Upton, J.P., Prakash, S. and Heideker, J. Nature 550.7677 (2017): 534.

Protective effect of in ovo treatment with the chicken cathelicidin analog D-CATH-2 against avian pathogenic E. coli.
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FAM标记说明:
Carboxyfluorescein (FAM) is fluorophor with an excitation at 492 nm ▉ and emission of 517 nm ▉. Donors like FAM and 5FAM are often paired together with acceptors (CPQ2) for FRET experiments.

FAM标记肽的相关文献:
Porous Silicon Nanoparticle Delivery of Tandem Peptide Anti-Infectives for the Treatment of Pseudomonas aeruginosa Lung Infections.
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Ultrasensitive tumor-penetrating nanosensors of protease activity.
Kwon, Ester J., Jaideep S. Dudani, and Sangeeta N. Bhatia. Nature Biomedical Engineering 1 (2017): 0054.

Seneca Valley Virus 3C pro Substrate Optimization Yields Efficient Substrates for Use in Peptide-Prodrug Therapy.
Miles, Linde A., et al. PloS One 10.6 (2015): e0129103.

The function of the milk-clotting enzymes bovine and camel chymosin studied by a fluorescence resonance energy transfer assay.
Jensen, Jesper Langholm, et al. Journal of Dairy Science 98.5 (2015): 2853-2860.

A comparison of modular PEG incorporation strategies for stabilization of peptide-siRNA nanocomplexes.
Lo, Justin H., et al. Bioconjugate Chemistry (2016).