Part:BBa_K4609050

DMBT1-sfGFP

DMBT1 protein is co-expressed with sfGFP protein, and the expression of DMBT1 protein can be visually checked by detecting the content of sfGFP protein. DMBT1 (Missing 1 in malignant brain tumors) is a approximately 340 kDa glycoprotein that is first isolated from saliva, belongs to the highly conserved scavorator receptor-rich cysteine (SRCR) protein superfamily, and is involved in mucosal innate immunity. From the n-terminal, DMBT1 contains 13 highly homologous SRCR domains separated by SIDs (SRCR diffusion domains). The 14th SRCR domain is separated from the other 13 by the CUB (C1r/C1s Uegf Bmp1) domain, where another domain separates the 14th domain from the ZP (Zpellucida) domain that forms the C-terminus. DMBT1 is expressed in a variety of tissues and body fluids and can undergo modifications that affect its function at specific sites. Due to alternative splicing and post-translational modification, different human DMBT1 alleles exist within the population, and different isotypes exist in various tissues. In salival-derived DMBT1SAG, about 25% of the molecular weight is due to glycosylation (about 10% for n-glycosylation and about 15% for O-glycosylation). DMBT1 protects mucosal tissue by inhibiting the twitching movement of Pseudomonas aeruginosa. Bacterial twitch movement is a surface-related movement commonly used by Gram-negative bacteria, driven by the extension and retraction of type IV pili (T4P). In Pseudomonas aeruginosa, T4P is a polymer composed mainly of PilA subunits. The extension and retraction required for movement are powered by the ATPases PilB, PilU and PilT. DMBT1 binds to Pseudomonas aeruginosa and inhibits twitch movement, a pile-mediated movement important for virulence. Binding of DMBT1 to multiple bacterial species involves its conserved clearance receptor cysteine-rich (SRCR) domain, localized to the 16-polymeric consensus sequence peptide SRCRP2. The inhibition of DMBT1 can be affected by temperature. Due to heat sensitivity, the thermal denaturated DMBT1 loses the ability to inhibit tic movement and shows a decrease in fimbrial binding (~40%). SRCR/SID components are required to inhibit the twitch movement of bacteria, and N-glycosylation of DMBT1 has a significant effect on inhibiting the twitch movement of Pseudomonas aeruginosa.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]