Part:BBa_K1080003
GUN4
Description & Function
File:Https://static.igem.org/mediawiki/2014/3/37/MQ-Gun4.png 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
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 719
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 404
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE 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
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