Difference between revisions of "Part:BBa K1080003"
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<br>GUN4 (Genomes uncoupled 4) is a protein which is found only in aerobic photosynthetic organisms. It is a tetrapyrrole-binding protein which controls levels of magnesium-protoporhyrin IX which is a crucial component for the production of chlorophyll within photosynthetic organisms. GUN4 is thought to participate in plastid-to-nucleus signalling by regulating magnesium-protoporphyrin IX synthesis or trafficking<br>
 
<br>GUN4 (Genomes uncoupled 4) is a protein which is found only in aerobic photosynthetic organisms. It is a tetrapyrrole-binding protein which controls levels of magnesium-protoporhyrin IX which is a crucial component for the production of chlorophyll within photosynthetic organisms. GUN4 is thought to participate in plastid-to-nucleus signalling by regulating magnesium-protoporphyrin IX synthesis or trafficking<br>
 
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<b><h4>Biobrick Design:</h4></b>
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Source Genbank accession: NW_001843510.1
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Source Uniprot reference: A8ILP2
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cDNA gene sequence from Chlamydomonas reinhardtii was sourced from NCBI database, chlorophyll targeting sequence was removed. EcoRI/XbaI/SpeI/PstI restriction sites were removed via codon adjustment, biobrick prefix and RBS were added to start of gene, biobrick suffix added to end of gene.
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<b>Biobrick construction:</b> Restriction and ligation of 1 synthesised DNA fragment into BB vector.
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Revision as of 02:33, 16 October 2014

GUN4


Description & Function


GUN4 (Genomes uncoupled 4) is a protein which is found only in aerobic photosynthetic organisms. It is a tetrapyrrole-binding protein which controls levels of magnesium-protoporhyrin IX which is a crucial component for the production of chlorophyll within photosynthetic organisms. GUN4 is thought to participate in plastid-to-nucleus signalling by regulating magnesium-protoporphyrin IX synthesis or trafficking

Biobrick Design:

Source Genbank accession: NW_001843510.1

Source Uniprot reference: A8ILP2

cDNA gene sequence from Chlamydomonas reinhardtii was sourced from NCBI database, chlorophyll targeting sequence was removed. EcoRI/XbaI/SpeI/PstI restriction sites were removed via codon adjustment, biobrick prefix and RBS were added to start of gene, biobrick suffix added to end of gene.

Biobrick construction: Restriction and ligation of 1 synthesised DNA fragment into BB vector.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 719
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 404
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]




Ribbon structure of protein

http://pfam.sanger.ac.uk/structure/getimage?id=1y6i


Amino acid sequence

MAMRVTVAAG KLDSVSLFGG DTASLMGGSQ TVEKKKSGKE AVMEVQLSST AGIDYTVLRD
HLANGEFREA EDETRALLIK LAGPEAVKRN WVYFTEVKNI SVTDFQTLDN LWKASSNNKF
GYSVQKEIWV QNQKRWPKFF KQIDWTQGEN NNYRKWPMEF IYSMDAPRGH LPLTNALRGT
QLFQAIMEHP AFEKSSTAKT LDQKAAEAAG RTQSLF

References and documentation are available. Please note the modified algorithm for extinction coefficient.


Number of amino acids: 216

Molecular weight: 24350.6

Theoretical pI: 8.98


Amino acid composition:

Ala(A) 21 ( 9.7% )
Arg(R) 10 ( 4.6% )
Asn(N) 11 ( 5.1% )
Asp(D) 10 ( 4.6% )
Cys(C) 00 ( 0.0% )
Gln(Q) 12 ( 5.6% )
Glu(E) 15 ( 6.9% )
Gly(G) 14 ( 6.5% )
His(H) 03 ( 1.4% )
Ile(I) 07 ( 3.2% )
Leu(L) 17 ( 7.9% )
Lys(K) 18 ( 8.3% )
Met(M) 07 ( 3.2% )
Phe(F) 11 ( 5.1% )
Pro(P) 06 ( 2.8% )
Ser(S) 15 ( 6.9% )
Thr(T) 15 ( 6.9% )
Trp(W) 06 ( 2.8% )
Tyr(Y) 05 ( 2.3% )
Val(V) 13 ( 6.0% )
Pyl(O) 00 ( 0.0% )
Sec(U) 00 ( 0.0% )

(B)   0	  0.0%
(Z)   0	  0.0%
(X)   0	  0.0%


Total number of negatively charged residues (Asp + Glu): 25 Total number of positively charged residues (Arg + Lys): 28


Atomic composition:

Carbon C 1085 Hydrogen H 1693 Nitrogen N 299 Oxygen O 325 Sulfur S 7

Formula: C1085H1693N299O325S7 Total number of atoms: 3409

Extinction coefficients:

Extinction coefficients are in units of M-1 cm-1, at 280 nm measured in water.

Ext. coefficient 40450 Abs 0.1% (=1 g/l) 1.661


Estimated half-life:

The N-terminal of the sequence considered is M (Met).

The estimated half-life is: 

                            30 hours (mammalian reticulocytes, in vitro).
                           >20 hours (yeast, in vivo).
                           >10 hours (Escherichia coli, in vivo).


Instability index:

The instability index (II) is computed to be 22.44 This classifies the protein as stable.


Aliphatic index: 70.51

Grand average of hydropathicity (GRAVY): -0.508


Source

Chlamydomonas reinhardtii

References

Brzezowski, P., et al. (2014). "The GUN4 protein plays a regulatory role in tetrapyrrole biosynthesis and chloroplast-to-nucleus signalling in Chlamydomonas reinhardtii." Plant J. 79(2): 285-98