Coding

Part:BBa_K4165079

Designed by: Mennatallah Mahmoud Mohamed Abdelzaher Turky   Group: iGEM22_CU_Egypt   (2022-09-30)
Revision as of 13:46, 6 October 2022 by Ahmedsameh (Talk | contribs)


SPINK2 (Serine Peptidase Inhibitor Kazal type 2).

This basic part encodes Human serine protease inhibitor known as SPINK2 which is able to inhibit trypsin-like proteases, like HtrA1 (BBa_K4165004).

Usage and Biology

This type of family encodes for a type of inhibitor that is able to inhibit trypsins. The main function of the inhibitor is hindering the premature activation of proacrosin and many other proteases, thus preventing defects in the spermiogenesis process. The inhibitor binds to trypsin proteases and since the catalytic core of HtrA1 (BBa_K4165004) is considered as a trypsin-like catalytic domain, so this inhibitor also is considered to inhibit the function of HtrA1 [1] - [3].

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 105
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 105
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 105
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 105
  • 1000
    COMPATIBLE WITH RFC[1000]


Functional Parameters

GC% Content 59.1% Isoelectric point (PI) 8.811 Charge at pH 7 5.702 Molecular Weight (Protein) 9.291 kDa

It has x-ray structure, NMR, and a predicted model (AlphaFold2). PDB structure

X-ray structure: https://www.rcsb.org/structure/2JXD Molprobity: Clash Score: Ramachandran Favoured: Ramachandran Outliers: Rotamers Outliers: C-beta Deviations: Q-Mean:

                 Figure 1.: A graphical illustration showing the structure of the inhibitor (X-ray).


NMR: https://www.rcsb.org/structure/2JXD Molprobity: Clash Score: Ramachandran Favoured: Ramachandran Outliers: Rotamers Outliers: C-beta Deviations: Q-Mean:


                 Figure 2.: A graphical illustration showing the structure of the inhibitor (NMR).


AlphaFold: https://alphafold.ebi.ac.uk/entry/P20155 Molprobity: Clash Score: Ramachandran Favoured: Ramachandran Outliers: Rotamers Outliers: C-beta Deviations: Q-Mean:


                 Figure 3.: A graphical illustration showing the structure of the inhibitor (AlphaFold).


References

1 - Frochaux, V., Hildebrand, D., Talke, A., Linscheid, M. W., & Schlüter, H. (2014). Alpha-1-antitrypsin: a novel human high temperature requirement protease A1 (HTRA1) substrate in human placental tissue. PloS one, 9(10), e109483. 2 - Grau, S., Baldi, A., Bussani, R., Tian, X., Stefanescu, R., Przybylski, M., ... & Ehrmann, M. (2005). Implications of the serine protease HtrA1 in amyloid precursor protein processing. Proceedings of the National Academy of Sciences, 102(17), 6021-6026. 3 - Eigenbrot, C., Ultsch, M., Lipari, M. T., Moran, P., Lin, S. J., Ganesan, R., ... & Kirchhofer, D. (2012). Structural and functional analysis of HtrA1 and its subdomains. Structure, 20(6), 1040-1050.

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