Difference between revisions of "Part:BBa K1080011"
 
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__NOTOC__
 
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<partinfo>BBa_K1080011 short</partinfo>
 
<partinfo>BBa_K1080011 short</partinfo>
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<br><b> Magnesium chelatase subunit I </b> - The second gene which catalyzes the insertion of magnesium ion into protoporphyrin IX to yield Mg-protoporphyrin IX. Forms an ATP dependent hexameric ring complex and a complex with the ChlD subunit.
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<br><b> Magnesium chelatase subunit I </b> - The second gene which catalyzes the insertion of magnesium ion into protoporphyrin IX to yield Mg-protoporphyrin IX. Forms an ATP dependent hexameric ring complex and a complex with the [https://parts.igem.org/Part:BBa_K1080002 ChlD] subunit.
 
<br>
 
<br>
 
===Usage and Biology===
 
===Usage and Biology===
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MPSTKAAKKP NFPFVKIQGQ EEMKLALLLN VVDPNIGGVL IMGDRGTAKS VAVRALVDML<br> PDIDVVEGDA FNSSPTDPKF MGPDTLQRFR NGEKLPTVRM RTPLVELPLG ATEDRICGTI<br> DIEKALTQGI KAYEPGLLAK ANRGILYVDE VNLLDDGLVD VVLDSSASGL NTVEREGVSI<br> VHPARFIMIG SGNPQEGELR PQLLDRFGMS VNVATLQDTK QRTQLVLDRL AYEADPDAFV<br> DSCKAEQTAL TDKLEAARQR LRSVKISEEL QILISDICSR LDVDGLRGDI VINRAAKALV<br> AFEGRTEVTT NDVERVISGC LNHRLRKDPL DPIDNGTKVA ILFKRMTDPE IMKREEEAKK<br> </FONT>
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MPS
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Please note the modified algorithm for extinction coefficient.
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--------------------------------------------------------------------------------
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Number of amino acids: 393
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Molecular weight: 42954.3
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Theoretical pI: 5.78
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Amino acid composition:
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<br>
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Ala (A)  38   9.7%<br>
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Arg (R)  30   7.6%<br>
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Asn (N)  14   3.6%<br>
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Asp (D)  32   8.1%<br>
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Cys (C)  4   1.0%<br>
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Gln (Q)  12   3.1%<br>
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Glu (E)  28   7.1%<br>
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Gly (G)  29   7.4%<br>
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His (H)  2   0.5%<br>
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Ile (I)  22   5.6%<br>
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Leu (L)  41 10.4%<br>
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Lys (K)  27   6.9%<br>
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Met (M)  10   2.5%<br>
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Phe (F)  10   2.5%<br>
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Pro (P)  22   5.6%<br>
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Ser (S)  16   4.1%<br>
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Thr (T)  21   5.3%<br>
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Trp (W)  1   0.3%<br>
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Tyr (Y)  3   0.8%<br>
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Val (V)  31   7.9%<br>
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Pyl (O)  0   0.0%<br>
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Sec (U)  0   0.0%<br>
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(B)  0   0.0%
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(Z)  0   0.0%
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(X)  0   0.0%
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Total number of negatively charged residues (Asp + Glu): 60
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Total number of positively charged residues (Arg + Lys): 57
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Atomic composition:
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Carbon      C       1875
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Hydrogen    H       3103
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Nitrogen    N       541
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Oxygen      O       580
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Sulfur      S         14
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Formula: C1875H3103N541O580S14
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Total number of atoms: 6113
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Extinction coefficients:
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Extinction coefficients are in units of  M-1 cm-1, at 280 nm measured in water.
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Ext. coefficient    10220
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Abs 0.1% (=1 g/l)  0.238, assuming all pairs of Cys residues form cystines
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Ext. coefficient    9970
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Abs 0.1% (=1 g/l)  0.232, assuming all Cys residues are reduced
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Estimated half-life:
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The N-terminal of the sequence considered is M (Met).
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The estimated half-life is:
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                            30 hours (mammalian reticulocytes, in vitro).
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                            >20 hours (yeast, in vivo).
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                            >10 hours (Escherichia coli, in vivo).
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Instability index:
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The instability index (II) is computed to be 32.87
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This classifies the protein as stable.
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Aliphatic index: 95.06
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Grand average of hydropathicity (GRAVY): -0.296
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===Source===
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<i>Chlamydomonas reinhardtii</i>
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<!-- Uncomment this to enable Functional Parameter display
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===Functional Parameters===
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<partinfo>BBa_K1080011 parameters</partinfo>
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<!-- -->
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Latest revision as of 10:01, 17 October 2014

ChlI2
Magnesium chelatase subunit I - The second gene which catalyzes the insertion of magnesium ion into protoporphyrin IX to yield Mg-protoporphyrin IX. Forms an ATP dependent hexameric ring complex and a complex with the ChlD subunit.

Usage and Biology

The chelation of magnesium in protoporphyrin IX is dependant on several genes. These include ChlI, ChlD, ChlH and GUN4. According to previous research, the ChlI component of this pathway proves essential in the chelation of magnesium. The ChlI gene encodes 2 isoforms, ChlI1 and ChlI2. The already familiar function of ChlI1 is well known and documented. The second isoform, ChlI2 is also integral in magnesium chelation complementing ChlI1 [http://www.plantphysiol.org/content/150/2/636.full]




ChlI catalysis reaction.png
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+.


Operon usage


PSB1C3 Operon 1.png


Figure 2: This gene has been used in an operon with other genes responsible for the synthesis pathway from Protoporphyrin IX to MG-protoporphyrin IX. ChlI1 and ChlD form an Mg-chelatase complex, which acts upon protoporphyrin, catalysing the insertion of a Magnesium ion into the centre of the protoporphyrin IX. GUN4 aids this reaction by activating the Mg-chelatase enzyme complex.
The plasmid is under the control of the lac promoter.
The entire operon can be found at BBa_K1326008.
Future plans involve adding ChlH to this operon, which binds Mg-chelatase with GUN4 to protoporphyrin.

ChlI structure

ChlI protein crystal structure.png
Figure 3: ChlI protein crystal structure.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 523
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]




Amino Acid Sequence

MPS