IGEM:MIT/2007/Ideas3
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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
- 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 |
- 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 |
- 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 |
- 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 |
- 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 |
- 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 |
- 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 |
- 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 |
- 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 |
- 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 |
- 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 |
- 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 |
- Samuelson P, Gunneriusson E, Nygren PA, and Ståhl S. Display of proteins on bacteria. J Biotechnol. 2002 Jun 26;96(2):129-54. DOI:10.1016/s0168-1656(02)00043-3 |
- 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 |
- 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.
- 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 |
- 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. DOI:10.1016/0378-1119(95)00058-e |
- 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 |
- Deming TJ. Mussel byssus and biomolecular materials. Curr Opin Chem Biol. 1999 Feb;3(1):100-5. DOI:10.1016/s1367-5931(99)80018-0 |
- Burzio LA and Waite JH. Cross-linking in adhesive quinoproteins: studies with model decapeptides. Biochemistry. 2000 Sep 12;39(36):11147-53. DOI:10.1021/bi0002434 |
- 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. DOI:10.1016/s0958-1669(97)80008-0 |
- 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. DOI:10.1016/0167-4838(86)90152-4 |
- 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.
- Waite JH and Qin X. Polyphosphoprotein from the adhesive pads of Mytilus edulis. Biochemistry. 2001 Mar 6;40(9):2887-93. DOI:10.1021/bi002718x |
- 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. DOI:10.1074/jbc.270.34.20183 |
Mgfp-5 Amino Acid Sequence
1 sseeykggyy pgntyhyhsg gsyhgsgyhg gykgkyygka kkyyykykns gkykylkkar 61 kyhrkgykky ygggss
/lue
6.20.07
KNOWN PEPTIDES/PROTEINS AND CHARACTERISTICS (please update!)
- 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?)
- fp-151
- Hwang, et al 2007
- fp-5 flanked by six decapeptide repeats from fp-1
- Obstacles
- Unknown fp-1 repeat
- Positives AGK
- Purifies more easily than fp-5
- Not harmful to cells
- 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
OmpX
- 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. DOI:10.1128/jb.177.3.799-804.1995 |
Sequence for CPX
- Total length is 558 bp + length of passenger peptide
- Native SS - 69 bp
MKKIACLSALAAVLAFTAGTSVA
- Embedded SfiI restriction site - 15 bp
GQSGQ
- Passenger Peptide
XXXXXXXXXXXXXX
- Sequence - 18 bp
GGQSGQ
- S54-F148 - 285 bp
SGDYNKNQYYGITAGPAYRINDWASIYGVVGVGYGKFQTTEYPTYKHDTSDYGFSYGAGLQFNPMENVALDFSYEQSRIRSVDVGTWIAGVGYRF
- Join native C and N - 12 bp
GGSG
- A1-S53 - 159 bp
ATSTVTGGYAQSDAQGQMNKMGGFNLKYRYEEDNSPLGVIGSFTYTEKSRTAS
Papers!
- 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. DOI:10.1128/AEM.66.3.1243-1248.2000 |
- 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. DOI:10.1128/AEM.65.3.1092-1098.1999 |
- 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. DOI:10.1128/AEM.64.10.4068-4072.1998 |
- 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. DOI:10.1128/jb.177.14.4121-4130.1995 |
- 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. DOI:10.1046/j.1365-2958.2000.01983.x |