Part:BBa_K2664004
trc-ChlI1
trc ChlI1
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1
Illegal BglII site found at 304
Illegal BglII site found at 960
Illegal BglII site found at 1155
Illegal BamHI site found at 352 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Usage and Biology
In the early stages of chlorophyll biosynthesis is the chelation of magnesium into the protoprophyrin-IX complex. The ATP hydrolysis by CHLI subunit of magnesium chelatase is essential for the insertion of Mg. CHLI1 is an isoform of CHLI that encodes the . It has the action of an ATPase and is reduced by thioredoxin. Magnesium chelatase is sensitive to thiol groups and it has been found that the insertion of Mg2+ increases as CHLI1 is reduced [1].
Figure 1: Reaction catalysed by Mg-chelatase, inserting a magnesium ion into Protoporphyrin IX to make Mg-protoporphyrin IX.
Reaction:
ATP + protoporphyrin IX + Mg2+ + H2O = ADP + phosphate + Mg-protoporphyrin IX + 2 H+.
Biobrick Design:
Source Genbank accession: [http://www.ncbi.nlm.nih.gov/nuccore/NW_001843537.1?report=genbank&from=707985&to=715043 NW_001843537.1]
Source Uniprot reference: [http://www.uniprot.org/uniprot/A8IMZ5 A8IMZ5]
cDNA gene sequence from Chlamydomonas reinhardtii was sourced from NCBI database, chloroplast 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. (added trc)
Biobrick construction: Gibson assembly of 2 synthesised DNA fragments into BB vector. (added trc)
Protein Structure
Figure 3: ChlI protein crystal structure.
Amino Acid Sequence
MAATEVKAAE GRTEKELGQA RPIFPFTAIV GQDEMKLALI LNVIDPKIGG VMIMGDRGTG KSTTIRALAD LLPEMQVVAN DPFNSDPTDP ELMSEEVRNR
VKAGEQLPVS SKKIPMVDLP LGATEDRVCG TIDIEKALTE GVKAFEPGLL AKANRGILYV DEVNLLDDHL VDVLLDSAAS GWNTVEREGI SISHPARFIL
VGSGNPEEGE LRPQLLDRFG MHAQIGTVKD PRLRVQIVSQ RSTFDENPAA FRKDYEAGQM ALTQRIVDAR KLLKQGEVNY DFRVKISQIC SDLNVDGIRG
DIVTNRAAKA LAAFEGRTEV TPEDIYRVIP LCLRHRLRKD PLAEIDDGDR VREIFKQVFG ME
References and documentation are available. Please note the modified algorithm for extinction coefficient.
Number of amino acids: 362
Molecular weight: 39952.7
Theoretical pI: 5.11
Amino acid composition:
Ala (A) 31 8.6%
Arg (R) 28 7.7%
Asn (N) 12 3.3%
Asp (D) 29 8.0%
Cys (C) 3 0.8%
Gln (Q) 13 3.6%
Glu (E) 29 8.0%
Gly (G) 28 7.7%
His (H) 4 1.1%
Ile (I) 25 6.9%
Leu (L) 34 9.4%
Lys (K) 19 5.2%
Met (M) 10 2.8%
Phe (F) 12 3.3%
Pro (P) 19 5.2%
Ser (S) 14 3.9%
Thr (T) 17 4.7%
Trp (W) 1 0.3%
Tyr (Y) 4 1.1%
Val (V) 30 8.3%
Pyl (O) 0 0.0%
Sec (U) 0 0.0%
(B) 0 0.0% (Z) 0 0.0% (X) 0 0.0%
Total number of negatively charged residues (Asp + Glu): 58
Total number of positively charged residues (Arg + Lys): 47
Atomic composition:
Carbon C 1752 Hydrogen H 2857 Nitrogen N 499 Oxygen O 539 Sulfur S 13
Formula: C1752H2857N499O539S13 Total number of atoms: 5660
Extinction coefficients:
Extinction coefficients are in units of M-1 cm-1, at 280 nm measured in water.
Ext. coefficient 11585 Abs 0.1% (=1 g/l) 0.290, assuming all pairs of Cys residues form cystines
Ext. coefficient 11460
Abs 0.1% (=1 g/l) 0.287, assuming all Cys residues are reduced
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 29.73 This classifies the protein as stable.
Aliphatic index: 96.16
Grand average of hydropathicity (GRAVY): -0.250
Source
Chlamydomonas reinhardtii
References
| None |