Difference between revisions of "IGEM:MIT/2007/Ideas3"

From OpenWetWare
Line 81: Line 81:
**Hwang, et al 2007
**Hwang, et al 2007
** fp-5 flanked by six decapeptide repeats from fp-1 AGK
** fp-5 flanked by six decapeptide repeats from fp-1 AGK
*** fp-1 sequence: Ala-Lys-Pro-Ser-Tyr-diHyp-Hyp-Thr-DOPA-Lys
*** [http://www.biolbull.org/cgi/reprint/190/2/213?view=long&pmid=8652732 | fp-1 sequence: Ala-Lys-Pro-Ser-Tyr-diHyp-Hyp-Thr-DOPA-Lys]
*** diHyp = 3,4 Dihydroxyproline; Hyp= 4 hydroxyproline  
*** diHyp = 3,4 Dihydroxyproline; Hyp= 4 hydroxyproline  
**Positives AGK
**Positives AGK

Revision as of 12:17, 20 June 2007


  • Put Links and Papers you find for another category here
  • Put your initials next to a link if you're in the process of analyzing it

Metal Pumps and Metal Specific Promoters

Mussel Protein

Expressing proteins on E coli cell membranes

Water Purification Methods

Metal Pumps and Metal Specific Promoters

Mussel Protein

  1. Hwang DS, Yoo HJ, Jun JH, Moon WK, and Cha HJ. Expression of functional recombinant mussel adhesive protein Mgfp-5 in Escherichia coli. Appl Environ Microbiol. 2004 Jun;70(6):3352-9. DOI:10.1128/AEM.70.6.3352-3359.2004 | PubMed ID:15184131 | HubMed [Hwang04]
  2. Hwang DS, Gim Y, and Cha HJ. Expression of functional recombinant mussel adhesive protein type 3A in Escherichia coli. Biotechnol Prog. 2005 May-Jun;21(3):965-70. DOI:10.1021/bp050014e | PubMed ID:15932281 | HubMed [Hwang05]
  3. Hwang DS, Gim Y, Yoo HJ, and Cha HJ. Practical recombinant hybrid mussel bioadhesive fp-151. Biomaterials. 2007 Aug;28(24):3560-8. DOI:10.1016/j.biomaterials.2007.04.039 | PubMed ID:17507090 | HubMed [Hwang07]
  4. Hwang DS, Gim Y, Kang DG, Kim YK, and Cha HJ. Recombinant mussel adhesive protein Mgfp-5 as cell adhesion biomaterial. J Biotechnol. 2007 Jan 20;127(4):727-35. DOI:10.1016/j.jbiotec.2006.08.005 | PubMed ID:16979252 | HubMed [Hwang07a]
  5. Lee H, Scherer NF, and Messersmith PB. Single-molecule mechanics of mussel adhesion. Proc Natl Acad Sci U S A. 2006 Aug 29;103(35):12999-3003. DOI:10.1073/pnas.0605552103 | PubMed ID:16920796 | HubMed [Lee06]
  6. Lin Q, Gourdon D, Sun C, Holten-Andersen N, Anderson TH, Waite JH, and Israelachvili JN. Adhesion mechanisms of the mussel foot proteins mfp-1 and mfp-3. Proc Natl Acad Sci U S A. 2007 Mar 6;104(10):3782-6. DOI:10.1073/pnas.0607852104 | PubMed ID:17360430 | HubMed [Lin07]
  7. Wang J, Liu C, Lu X, and Yin M. Co-polypeptides of 3,4-dihydroxyphenylalanine and L-lysine to mimic marine adhesive protein. Biomaterials. 2007 Aug;28(23):3456-68. DOI:10.1016/j.biomaterials.2007.04.009 | PubMed ID:17475323 | HubMed [Wang07]
  8. Rice JJ, Schohn A, Bessette PH, Boulware KT, and Daugherty PS. Bacterial display using circularly permuted outer membrane protein OmpX yields high affinity peptide ligands. Protein Sci. 2006 Apr;15(4):825-36. DOI:10.1110/ps.051897806 | PubMed ID:16600968 | HubMed [Rice07]
  9. Dane KY, Chan LA, Rice JJ, and Daugherty PS. Isolation of cell specific peptide ligands using fluorescent bacterial display libraries. J Immunol Methods. 2006 Feb 20;309(1-2):120-9. DOI:10.1016/j.jim.2005.11.021 | PubMed ID:16448666 | HubMed [Dane06]
  10. Bessette PH, Rice JJ, and Daugherty PS. Rapid isolation of high-affinity protein binding peptides using bacterial display. Protein Eng Des Sel. 2004 Oct;17(10):731-9. DOI:10.1093/protein/gzh084 | PubMed ID:15531628 | HubMed [Bessette04]
  11. Etz H, Minh DB, Schellack C, Nagy E, and Meinke A. Bacterial phage receptors, versatile tools for display of polypeptides on the cell surface. J Bacteriol. 2001 Dec;183(23):6924-35. DOI:10.1128/JB.183.23.6924-6935.2001 | PubMed ID:11698382 | HubMed [Etz01]
  12. Hall SS, Mitragotri S, and Daugherty PS. Identification of peptide ligands facilitating nanoparticle attachment to erythrocytes. Biotechnol Prog. 2007 May-Jun;23(3):749-54. DOI:10.1021/bp060333l | PubMed ID:17469847 | HubMed [Hall07]
  13. Samuelson P, Gunneriusson E, Nygren PA, and Ståhl S. Display of proteins on bacteria. J Biotechnol. 2002 Jun 26;96(2):129-54. PubMed ID:12039531 | HubMed [Samuelson02]
  14. Wernérus H and Ståhl S. Biotechnological applications for surface-engineered bacteria. Biotechnol Appl Biochem. 2004 Dec;40(Pt 3):209-28. DOI:10.1042/BA20040014 | PubMed ID:15035661 | HubMed [Wernerus04]
  15. Gebhardt K, Lauvrak V, Babaie E, Eijsink V, and Lindqvist BH. Adhesive peptides selected by phage display: characterization, applications and similarities with fibrinogen. Pept Res. 1996 Nov-Dec;9(6):269-78. PubMed ID:9048419 | HubMed [Gebhart96]
  16. Kenan DJ, Walsh EB, Meyers SR, O'Toole GA, Carruthers EG, Lee WK, Zauscher S, Prata CA, and Grinstaff MW. Peptide-PEG amphiphiles as cytophobic coatings for mammalian and bacterial cells. Chem Biol. 2006 Jul;13(7):695-700. DOI:10.1016/j.chembiol.2006.06.013 | PubMed ID:16873017 | HubMed [Kenan06]
  17. Adey NB, Mataragnon AH, Rider JE, Carter JM, and Kay BK. Characterization of phage that bind plastic from phage-displayed random peptide libraries. Gene. 1995 Apr 14;156(1):27-31. PubMed ID:7737512 | HubMed [Adey95]
  18. Menendez A and Scott JK. The nature of target-unrelated peptides recovered in the screening of phage-displayed random peptide libraries with antibodies. Anal Biochem. 2005 Jan 15;336(2):145-57. DOI:10.1016/j.ab.2004.09.048 | PubMed ID:15620878 | HubMed [Menendez05]
  19. Deming TJ. Mussel byssus and biomolecular materials. Curr Opin Chem Biol. 1999 Feb;3(1):100-5. PubMed ID:10021411 | HubMed [Deming99]
  20. Burzio LA and Waite JH. Cross-linking in adhesive quinoproteins: studies with model decapeptides. Biochemistry. 2000 Sep 12;39(36):11147-53. PubMed ID:10998254 | HubMed [Burzio00]
  21. Burzio LO, Burzio VA, Silva T, Burzio LA, and Pardo J. Environmental bioadhesion: themes and applications. Curr Opin Biotechnol. 1997 Jun;8(3):309-12. PubMed ID:9206011 | HubMed [Burzio97]
  22. Marumo K and Waite JH. Optimization of hydroxylation of tyrosine and tyrosine-containing peptides by mushroom tyrosinase. Biochim Biophys Acta. 1986 Jul 25;872(1-2):98-103. PubMed ID:3089286 | HubMed [Marumo86]
  23. Waite JH. Evidence for a repeating 3,4-dihydroxyphenylalanine- and hydroxyproline-containing decapeptide in the adhesive protein of the mussel, Mytilus edulis L. J Biol Chem. 1983 Mar 10;258(5):2911-5. PubMed ID:6298211 | HubMed [Waite83]
  24. Waite JH and Qin X. Polyphosphoprotein from the adhesive pads of Mytilus edulis. Biochemistry. 2001 Mar 6;40(9):2887-93. PubMed ID:11258900 | HubMed [Waite01]
  25. Papov VV, Diamond TV, Biemann K, and Waite JH. Hydroxyarginine-containing polyphenolic proteins in the adhesive plaques of the marine mussel Mytilus edulis. J Biol Chem. 1995 Aug 25;270(34):20183-92. PubMed ID:7650037 | HubMed [Papov95]

All Medline abstracts: PubMed | HubMed

Mgfp-5 Amino Acid Sequence

 1 sseeykggyy pgntyhyhsg gsyhgsgyhg gykgkyygka kkyyykykns gkykylkkar
61 kyhrkgykky ygggss




  • Name of peptide/protein
    • Paper/Reference
    • Length in amino acids
    • Obstacles for synthesis?
    • Relevant characteristics/Other pertinent information

+ + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + +

  • Mgfp-5/Mefp-5
    • Hwang, et al
    • 75 aa
    • Obstacles AGK
      • "However, these attempts have failed to express functional and economical mussel adhesive proteins for several reasons, including a highly biased amino acid composition (5 amino acid types comprised �89% of the total amino acids), different codon usage preference between mussel and other expression systems (tRNA utilization problem), and small amount of adhesive produced" - Hwang
      • Presence of fp-5 has adverse effects on cell growth.
    • Positives AGK
      • Known sequence
      • Very adhesive
      • Has a 30% DOPA level, which no other foot protein comes close to
      • From M. galloprovincialis; Very adhesive; Low purification yield (but this last fact may not matter for us?)
  • Mgfp-1
    • Hwang, et al
    • … aa
  • Mgfp-3
    • Hwang, et al
    • … aa

  • Mefp-1
    • Hwang, et al
    • 10 aa ?
    • Not very adhesive
  • Mefp-5
  • Mefp-1

Water Purification Methods


  1. Mecsas J, Welch R, Erickson JW, and Gross CA. Identification and characterization of an outer membrane protein, OmpX, in Escherichia coli that is homologous to a family of outer membrane proteins including Ail of Yersinia enterocolitica. J Bacteriol. 1995 Feb;177(3):799-804. PubMed ID:7836315 | HubMed [Mecsas95]

[OmpX sequence]

Sequence for CPX

  • Total length is 558 bp + length of passenger peptide
  1. Native SS - 69 bp
  2. Embedded SfiI restriction site - 15 bp
  3. Passenger Peptide
  4. Sequence - 18 bp
  5. S54-F148 - 285 bp
  6. Join native C and N - 12 bp
  7. A1-S53 - 159 bp


  1. Samuelson P, Wernérus H, Svedberg M, and Ståhl S. Staphylococcal surface display of metal-binding polyhistidyl peptides. Appl Environ Microbiol. 2000 Mar;66(3):1243-8. PubMed ID:10698802 | HubMed [Samuelson00]
  2. Kotrba P, Dolecková L, de Lorenzo V, and Ruml T. Enhanced bioaccumulation of heavy metal ions by bacterial cells due to surface display of short metal binding peptides. Appl Environ Microbiol. 1999 Mar;65(3):1092-8. PubMed ID:10049868 | HubMed [Kotrba99]
  3. Pazirandeh M, Wells BM, and Ryan RL. Development of bacterium-based heavy metal biosorbents: enhanced uptake of cadmium and mercury by Escherichia coli expressing a metal binding motif. Appl Environ Microbiol. 1998 Oct;64(10):4068-72. PubMed ID:9758845 | HubMed [Biosorbants]
  4. Guzman LM, Belin D, Carson MJ, and Beckwith J. Tight regulation, modulation, and high-level expression by vectors containing the arabinose PBAD promoter. J Bacteriol. 1995 Jul;177(14):4121-30. PubMed ID:7608087 | HubMed [pBAD33]
  5. Koebnik R, Locher KP, and Van Gelder P. Structure and function of bacterial outer membrane proteins: barrels in a nutshell. Mol Microbiol. 2000 Jul;37(2):239-53. PubMed ID:10931321 | HubMed [koebnik00]

All Medline abstracts: PubMed | HubMed