Alignments for a candidate for tpiA in Methanosarcina mazei Go1

GapMind for Amino acid biosynthesis

Alignments for a candidate for tpiA in Methanosarcina mazei Go1

Align triose-phosphate isomerase (EC 5.3.1.1) (characterized)
to candidate WP_011033225.1 MM_RS06610 triose-phosphate isomerase

Query= BRENDA::Q12UK2
         (221 letters)



>NCBI__GCF_000007065.1:WP_011033225.1
          Length = 222

 Score =  335 bits (858), Expect = 5e-97
 Identities = 168/219 (76%), Positives = 193/219 (88%)

Query: 3   PLIVLNLKTYLEGTGEGAVRIARACKEVGEASGIEIAIAPQFCDIYRVASQVDVPVYSQH 62
           P I+LN KTY++GTG+GAV IA+ACK V E SGIEIA+APQ  DIYRVAS+V++ V+SQH
Sbjct: 4   PFILLNYKTYIQGTGKGAVDIAKACKTVSEESGIEIAVAPQLPDIYRVASEVELSVFSQH 63

Query: 63  LDGVGAGSFTGHAFAKCIKDAGAVGTLINHSECRLKLADIEASVTAAKGEGLRTIICTNN 122
           +DG+GAGSFTGH F KCIK+AGA GTLINHSE RL LA+IEAS+ AAK  GLR IICTNN
Sbjct: 64  MDGIGAGSFTGHVFGKCIKEAGAFGTLINHSERRLTLAEIEASLKAAKEFGLRAIICTNN 123

Query: 123 IATTAAAAALGPDYVAVEPPELIGSGIPVSKADPEVVTGSVAAVERIDPAVKVLCGAGIS 182
           + TTAAAAAL PDYVA+EPPELIGSGIPVSKADPEVV+GSV AV +I+P VKVLCGAGIS
Sbjct: 124 VPTTAAAAALVPDYVAIEPPELIGSGIPVSKADPEVVSGSVEAVSKINPDVKVLCGAGIS 183

Query: 183 KGEDLKAAIELGSVGVLLASGIVKAKDPKAALEDLVSLI 221
           KGEDL+AA++LGS GVLLASGIVKA DPKAALEDL+ L+
Sbjct: 184 KGEDLRAALDLGSQGVLLASGIVKASDPKAALEDLIRLV 222


Lambda     K      H
   0.316    0.134    0.378 

Gapped
Lambda     K      H
   0.267   0.0410    0.140 


Matrix: BLOSUM62
Gap Penalties: Existence: 11, Extension: 1
Number of Sequences: 1
Number of Hits to DB: 251
Number of extensions: 15
Number of successful extensions: 3
Number of sequences better than 1.0e-02: 1
Number of HSP's gapped: 1
Number of HSP's successfully gapped: 1
Length of query: 221
Length of database: 222
Length adjustment: 22
Effective length of query: 199
Effective length of database: 200
Effective search space:    39800
Effective search space used:    39800
Neighboring words threshold: 11
Window for multiple hits: 40
X1: 16 ( 7.3 bits)
X2: 38 (14.6 bits)
X3: 64 (24.7 bits)
S1: 41 (21.6 bits)
S2: 45 (21.9 bits)

Align candidate WP_011033225.1 MM_RS06610 (triose-phosphate isomerase)
to HMM TIGR00419 (tpiA: triose-phosphate isomerase (EC 5.3.1.1))

# hmmsearch :: search profile(s) against a sequence database
# HMMER 3.3.1 (Jul 2020); http://hmmer.org/
# Copyright (C) 2020 Howard Hughes Medical Institute.
# Freely distributed under the BSD open source license.
# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
# query HMM file:                  ../tmp/path.aa/TIGR00419.hmm
# target sequence database:        /tmp/gapView.4187528.genome.faa
# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -

Query:       TIGR00419  [M=228]
Accession:   TIGR00419
Description: tim: triose-phosphate isomerase
Scores for complete sequences (score includes all domains):
   --- full sequence ---   --- best 1 domain ---    -#dom-
    E-value  score  bias    E-value  score  bias    exp  N  Sequence                             Description
    ------- ------ -----    ------- ------ -----   ---- --  --------                             -----------
    3.3e-69  219.2   1.0    3.8e-69  219.0   1.0    1.0  1  NCBI__GCF_000007065.1:WP_011033225.1  


Domain annotation for each sequence (and alignments):
>> NCBI__GCF_000007065.1:WP_011033225.1  
   #    score  bias  c-Evalue  i-Evalue hmmfrom  hmm to    alifrom  ali to    envfrom  env to     acc
 ---   ------ ----- --------- --------- ------- -------    ------- -------    ------- -------    ----
   1 !  219.0   1.0   3.8e-69   3.8e-69       1     227 [.       5     208 ..       5     209 .. 0.99

  Alignments for each domain:
  == domain 1  score: 219.0 bits;  conditional E-value: 3.8e-69
                             TIGR00419   1 lviinfK.lnesvgkvelevaklaeevaseagvevavappfvdldvvkdeveseiqvaAqnvdavksGaftGe 72 
                                           ++++n+K + +++gk +  +ak ++ v++e+g+e+avap+  d+ +v+ eve  ++v  q++d + +G+ftG+
  NCBI__GCF_000007065.1:WP_011033225.1   5 FILLNYKtYIQGTGKGAVDIAKACKTVSEESGIEIAVAPQLPDIYRVASEVE--LSVFSQHMDGIGAGSFTGH 75 
                                           6899************************************************..******************* PP

                             TIGR00419  73 isAemlkdlGakgvligHsErRsllkeadeliekkvarlkelglksvvCvgetleereaartinnvattaaaa 145
                                           +  +++k++Ga g+li+HsErR  l +a+  ie+ +  +ke gl++++C+             nnv ttaaaa
  NCBI__GCF_000007065.1:WP_011033225.1  76 VFGKCIKEAGAFGTLINHSERR--LTLAE--IEASLKAAKEFGLRAIICT-------------NNVPTTAAAA 131
                                           **********************..*****..*******************.............********** PP

                             TIGR00419 146 AlepdvvAvEPveliGtGkpvskAeaevveksvrdhlkkvskevaesvrvlyGasvtaaedaelaaqldvdGv 218
                                           Al pd+vA+EP+eliG+G+pvskA++evv +sv+     ++ +++++v+vl+Ga+++++ed ++a++l+ +Gv
  NCBI__GCF_000007065.1:WP_011033225.1 132 ALVPDYVAIEPPELIGSGIPVSKADPEVVSGSVE-----AVSKINPDVKVLCGAGISKGEDLRAALDLGSQGV 199
                                           **********************************.....********************************** PP

                             TIGR00419 219 Llasavlka 227
                                           Llas+++ka
  NCBI__GCF_000007065.1:WP_011033225.1 200 LLASGIVKA 208
                                           ********9 PP



Internal pipeline statistics summary:
-------------------------------------
Query model(s):                            1  (228 nodes)
Target sequences:                          1  (222 residues searched)
Passed MSV filter:                         1  (1); expected 0.0 (0.02)
Passed bias filter:                        1  (1); expected 0.0 (0.02)
Passed Vit filter:                         1  (1); expected 0.0 (0.001)
Passed Fwd filter:                         1  (1); expected 0.0 (1e-05)
Initial search space (Z):                  1  [actual number of targets]
Domain search space  (domZ):               1  [number of targets reported over threshold]
# CPU time: 0.00u 0.00s 00:00:00.00 Elapsed: 00:00:00.00
# Mc/sec: 21.89
//
[ok]

This GapMind analysis is from Jul 25 2024. The underlying query database was built on Jul 25 2024.

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About GapMind

Each pathway is defined by a set of rules based on individual steps or genes. Candidates for each step are identified by using ublast (a fast alternative to protein BLAST) against a database of manually-curated proteins (most of which are experimentally characterized) or by using HMMer with enzyme models (usually from TIGRFam). Ublast hits may be split across two different proteins.

A candidate for a step is "high confidence" if either:

ublast finds a hit to a characterized protein at above 40% identity and 80% coverage, and bits >= other bits+10.
- (Hits to curated proteins without experimental data as to their function are never considered high confidence.)
HMMer finds a hit with 80% coverage of the model, and either other identity < 40 or other coverage < 0.75.

where "other" refers to the best ublast hit to a sequence that is not annotated as performing this step (and is not "ignored").

Otherwise, a candidate is "medium confidence" if either:

ublast finds a hit at above 40% identity and 70% coverage (ignoring otherBits).
ublast finds a hit at above 30% identity and 80% coverage, and bits >= other bits.
HMMer finds a hit (regardless of coverage or other bits).

Other blast hits with at least 50% coverage are "low confidence."

Steps with no high- or medium-confidence candidates may be considered "gaps." For the typical bacterium that can make all 20 amino acids, there are 1-2 gaps in amino acid biosynthesis pathways. For diverse bacteria and archaea that can utilize a carbon source, there is a complete high-confidence catabolic pathway (including a transporter) just 38% of the time, and there is a complete medium-confidence pathway 63% of the time. Gaps may be due to:

our ignorance of proteins' functions,
omissions in the gene models,
frame-shift errors in the genome sequence, or
the organism lacks the pathway.

GapMind relies on the predicted proteins in the genome and does not search the six-frame translation. In most cases, you can search the six-frame translation by clicking on links to Curated BLAST for each step definition (in the per-step page).

For more information, see:

the paper from 2019 on GapMind for amino acid biosynthesis
the paper from 2022 on GapMind for carbon sources
the source code
instructions for running GapMind on your computer
changes to Amino acid biosynthesis since the publication.

If you notice any errors or omissions in the step descriptions, or any questionable results, please let us know

by Morgan Price, Arkin group, Lawrence Berkeley National Laboratory

GapMind for Amino acid biosynthesis