Berkeley Structural Genomics Center

Target info for 1151B

This target is MP488 / MPN348, gi number 1674179 from Mycoplasma pneumoniae.

This gene has the annotation "5-formyl tetrahydrofolate cyclo-ligase (HI0858) homolog; similar to GenBank Accession Number D64160, from H. influenzae".

The target was selected as part of JMC's first set.

Available sections:


Experimental status of 1151B

Note: Dates prior to 10/24/01 may be inaccurate, due to conversion from a record-keeping system which tracked only the last experimental date, to a new system which keeps track of all dates. The user is the person who entered the data, not the person who did the experiment. This will also change in the future.

ExperimentResultUserExperiment Date
SelectedDonejmc2001-08-28
ClonedDoner_kim2002-06-20
Expression testedDoner_kim2002-06-20
Solubility testedSr_kim2002-06-20
PurifiedDonedewemmer2002-09-05
CrystallizedDonedhshin2002-12-16
Diffraction quality crystalsDonedhshin2002-12-16
Native diffraction dataDonedhshin2002-12-16
Phasing diffraction dataDonedhshin2002-12-16
Traceable mapDonedhshin2002-12-16
Crystal structureDonedhshin2004-08-05
NMR characterizationDonedewemmer2002-09-05
HSQCDonedewemmer2002-09-05
In PDB1SBQ 1U3F 1U3Gdhshin2004-08-05
Suggested work stop19182 19182dallen2003-05-20

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Potential MP Homologues of 1151B

MP genelog10(PSIBLAST E)% identical% coverage (of MP gene)
MP488 / MPN348-46.40100.00100.00

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1151B DNA sequence

ATGGACAAAAATGCCTTAAGAAAACAAATTCTGCAAAAAAGAATGGCATTAAGTACCATTGAAAAAAGTCACTTAGATCA
AAAGATTAATCAAAAATTAGTTGCTTTTTTAACTCCTAAGCCATGCATTAAAACAATTGCACTTTATGAACCCATTAAAA
ATGAGGTTACTTTTGTTGACTTCTTCTTTGAGTTTTTAAAGATTAACCAAATAAGAGCTGTTTACCCCAAGGTAATAAGT
GATACCGAAATTATCTTTATTGATCAGGAGACAAATACATTCGAACCTAATCAAATAGACTGCTTCCTAATACCATTGGT
AGGCTTTAATAAAGACAATTACCGTCTAGGCTTTGGCAAGGGCTATTATGACCGTTATTTAATGCAATTAACTAGACAAC
AACCTAAAATAGGAATAGCGTACAGTTTCCAAAAAGGTGATTTTTTAGCCGATCCTTGGGATGTACAACTAGACTTAATT
ATTAATGATGAATAG

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1151B Codon Usage

(165 codons)
fields: [triplet] [frequency: per thousand] ([number])

UUU48.5(   8)  UCU0.0(   0)  UAU24.2(   4)  UGU0.0(   0)  
UUC30.3(   5)  UCC0.0(   0)  UAC18.2(   3)  UGC12.1(   2)  
UUA60.6(  10)  UCA0.0(   0)  UAA0.0(   0)  UGA0.0(   0)  
UUG6.1(   1)  UCG0.0(   0)  UAG6.1(   1)  UGG6.1(   1)  

CUU6.1(   1)  CCU24.2(   4)  CAU0.0(   0)  CGU12.1(   2)  
CUC0.0(   0)  CCC12.1(   2)  CAC6.1(   1)  CGC0.0(   0)  
CUA18.2(   3)  CCA12.1(   2)  CAA66.7(  11)  CGA0.0(   0)  
CUG6.1(   1)  CCG0.0(   0)  CAG6.1(   1)  CGG0.0(   0)  

AUU66.7(  11)  ACU18.2(   3)  AAU48.5(   8)  AGU24.2(   4)  
AUC6.1(   1)  ACC12.1(   2)  AAC6.1(   1)  AGC0.0(   0)  
AUA36.4(   6)  ACA18.2(   3)  AAA60.6(  10)  AGA24.2(   4)  
AUG18.2(   3)  ACG0.0(   0)  AAG30.3(   5)  AGG0.0(   0)  

GUU24.2(   4)  GCU12.1(   2)  GAU42.4(   7)  GGU6.1(   1)  
GUC0.0(   0)  GCC12.1(   2)  GAC36.4(   6)  GGC24.2(   4)  
GUA18.2(   3)  GCA12.1(   2)  GAA30.3(   5)  GGA6.1(   1)  
GUG0.0(   0)  GCG6.1(   1)  GAG18.2(   3)  GGG0.0(   0)  

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1151B Potential Cloning Problems

No UGA codons.
1 restriction site(s) found.

Details (problem DNA region is in upper case, with the key on the line below):

Key:

atggacaaaaatgccttaagaaaacaaattctgcaaaaaagaatggcattaagtaccatt
                                                            
gaaaaaagtcacttagatcaaaagattaatcaaaaattagttgcttttttaactcctaag
                                                            
ccatgcattaaaacaattgcactttatgaacccattaaaaatgaggttacttttgttgac
                                                            
ttcttctttgagtttttaaagattaaccaaataagagctgtttaccccaaggtaataagt
                                                            
gataccgaaattatctttatTGATCAggagacaaatacattcgaacctaatcaaatagac
                    3                                       
tgcttcctaataccattggtaggctttaataaagacaattaccgtctaggctttggcaag
                                                            
ggctattatgaccgttatttaatgcaattaactagacaacaacctaaaataggaatagcg
                                                            
tacagtttccaaaaaggtgattttttagccgatccttgggatgtacaactagacttaatt
                                                            
attaatgatgaatag
               

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1151B Automatically Selected Primers

Note: These were generated using Hisao's algorithm, and do not take into account possible mispriming, self-complementarity, or requirements for mutations anywhere in the sequences. The -R1 primers are for the N-terminal end, and begin with an NdeI restriction site (CATATG). The -R2 primers are for the C-terminal end, and begin with a BamHI site (GGATCC) if the target has no other BamHI sites. If the target does have a BamHI site, the -R2 primer wil begin with a BglII site (AGATCT), unless the target has no BglII sites. If the target has both sites, the design program gives up and doesn't prepend a restriction site to the -R2 primer!

Predicted Tm was obtained using the Tm determination tool at the Virtual Genome Center.

Primer IDPredicted Tmsequence
1151B-R160.10CATATGGACAAAAATGCCTTAAGAAAA
1151B-R260.80GGATCCTTATTCATCATTAATAATTAAGTCTAGTTGTACATCC
1151B-R360.80GGATCCTTATTCATCATTAATAATTAAGTCTAGTTGTACATCC
1151B-R1063.00GGCGGTGGTGGCGGCATGGACAAAAATGCCTTAAGAAAACA
1151B-R1163.40GTTCTTCTCCTTTGCGCCCCTATTCATCATTAATAATTAAGTCTAGTTGTACATCCC

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1151B Translation

Note: This was generated using the standard codon usage table; UGA codons in MP/MP genes will show up as terminators ("*")

atggacaaaaatgccttaagaaaacaaattctgcaaaaaagaatggcattaagtaccatt
 M  D  K  N  A  L  R  K  Q  I  L  Q  K  R  M  A  L  S  T  I 
gaaaaaagtcacttagatcaaaagattaatcaaaaattagttgcttttttaactcctaag
 E  K  S  H  L  D  Q  K  I  N  Q  K  L  V  A  F  L  T  P  K 
ccatgcattaaaacaattgcactttatgaacccattaaaaatgaggttacttttgttgac
 P  C  I  K  T  I  A  L  Y  E  P  I  K  N  E  V  T  F  V  D 
ttcttctttgagtttttaaagattaaccaaataagagctgtttaccccaaggtaataagt
 F  F  F  E  F  L  K  I  N  Q  I  R  A  V  Y  P  K  V  I  S 
gataccgaaattatctttattgatcaggagacaaatacattcgaacctaatcaaatagac
 D  T  E  I  I  F  I  D  Q  E  T  N  T  F  E  P  N  Q  I  D 
tgcttcctaataccattggtaggctttaataaagacaattaccgtctaggctttggcaag
 C  F  L  I  P  L  V  G  F  N  K  D  N  Y  R  L  G  F  G  K 
ggctattatgaccgttatttaatgcaattaactagacaacaacctaaaataggaatagcg
 G  Y  Y  D  R  Y  L  M  Q  L  T  R  Q  Q  P  K  I  G  I  A 
tacagtttccaaaaaggtgattttttagccgatccttgggatgtacaactagacttaatt
 Y  S  F  Q  K  G  D  F  L  A  D  P  W  D  V  Q  L  D  L  I 
attaatgatgaatag
 I  N  D  E  * 

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1151B AA Sequence

MDKNALRKQILQKRMALSTIEKSHLDQKINQKLVAFLTPKPCIKTIALYEPIKNEVTFVDFFFEFLKINQIRAVYPKVIS
DTEIIFIDQETNTFEPNQIDCFLIPLVGFNKDNYRLGFGKGYYDRYLMQLTRQQPKIGIAYSFQKGDFLADPWDVQLDLI
INDE

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1151B Protein Parameters

This information was obtained using the Protein Parameters tool on the ExPASy Molecular Biology Server.

Number of amino acids: 164

Molecular weight: 19265.3

Theoretical pI: 6.60

Amino acid composition:

Ala (A)   7	  4.3%
Arg (R)   6	  3.7%
Asn (N)   9	  5.5%
Asp (D)  13	  7.9%
Cys (C)   2	  1.2%
Gln (Q)  12	  7.3%
Glu (E)   8	  4.9%
Gly (G)   6	  3.7%
His (H)   1	  0.6%
Ile (I)  18	 11.0%
Leu (L)  16	  9.8%
Lys (K)  15	  9.1%
Met (M)   3	  1.8%
Phe (F)  13	  7.9%
Pro (P)   8	  4.9%
Ser (S)   4	  2.4%
Thr (T)   8	  4.9%
Trp (W)   1	  0.6%
Tyr (Y)   7	  4.3%
Val (V)   7	  4.3%

Asx (B)   0	  0.0%
Glx (Z)   0	  0.0%
Xaa (X)   0	  0.0%

Total number of negatively charged residues (Asp + Glu): 21
Total number of positively charged residues (Arg + Lys): 21

Atomic composition:

Carbon      C	       888
Hydrogen    H	      1381
Nitrogen    N	       221
Oxygen      O	       247
Sulfur      S	         5

Formula: C888H1381N221O247S5
Total number of atoms: 2742

Extinction coefficients:

Conditions: 6.0 M guanidium hydrochloride
            0.02 M phosphate buffer
            pH 6.5

Extinction coefficients are in units of  M-1 cm-1 .

The first table lists values computed assuming ALL Cys 
residues appear as half cystines, whereas the second table 
assumes that NONE do. 

                      276     278     279     280     282
                       nm      nm      nm      nm      nm
Ext. coefficient    15695   15527   15195   14770   14120
Abs 0.1% (=1 g/l)   0.815   0.806   0.789   0.767   0.733



                      276     278     279     280     282
                       nm      nm      nm      nm      nm
Ext. coefficient    15550   15400   15075   14650   14000
Abs 0.1% (=1 g/l)   0.807   0.799   0.782   0.760   0.727


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 17.46
This classifies the protein as stable.



Aliphatic index: 97.50

Grand average of hydropathicity (GRAVY): -0.237

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