Alignments for a candidate for trpA in Pseudomonas fluorescens FW300-N2E2

GapMind for Amino acid biosynthesis

Alignments for a candidate for trpA in Pseudomonas fluorescens FW300-N2E2

Align Tryptophan synthase alpha chain; EC 4.2.1.20 (characterized, see rationale)
to candidate Pf6N2E2_4047 Tryptophan synthase alpha chain (EC 4.2.1.20)

Query= uniprot:H0BV16
         (269 letters)



>FitnessBrowser__pseudo6_N2E2:Pf6N2E2_4047
          Length = 270

 Score =  227 bits (578), Expect = 2e-64
 Identities = 122/271 (45%), Positives = 171/271 (63%), Gaps = 9/271 (3%)

Query: 1   MSRIASTFSALQAQGRKALIPYVTAGFPFADITPALMHGMVEAGADVIELGVPFSDPMAD 60
           MSR+ + F+ L+ Q R AL+ +VTAG P  D + A++ G+  AGADVIELG+PF+DPMAD
Sbjct: 1   MSRLQTRFAELKEQNRAALVTFVTAGDPDYDTSLAILKGLPAAGADVIELGMPFTDPMAD 60

Query: 61  GPVIQKAGEKALSLGIGMVQVLDHVREFRKRNNTTPVVLMGYANPVERYDQKHGEGAFVR 120
           GP IQ A  +AL     + + L  VREFRK N+ TP+VLMGY NP+ RY    G   F+ 
Sbjct: 61  GPAIQLANIRALGAQQNLAKTLQMVREFRKDNSETPLVLMGYFNPIHRY----GVQRFIG 116

Query: 121 DSAAAGVDGVLIVDYPPEECEAFAASLRAHGMDLIFLLAPTSTDERMAEVARVASGYVYY 180
           ++  +GVDG+++VD PPE         +A G+D I L  PT+ D R+  V   +SG+VYY
Sbjct: 117 EALESGVDGLIVVDMPPEHNSELCEPAQAAGLDFIRLTTPTTDDVRLPTVLNGSSGFVYY 176

Query: 181 VSLKGVTGSGALDTAAVEQMLPRIRQHVTIPVGVGFGIRDAATAQAIGKVADAVVIGSRI 240
           VS+ GVTG+GA     VE+ + R+R+H  +P+ +GFGIR    A AI ++AD VV+GS +
Sbjct: 177 VSVAGVTGAGAATLEHVEEAVARLRRHTDLPISIGFGIRTPEQAAAIARLADGVVVGSAL 236

Query: 241 IQLIEDQEHAK-----VVPLTIDFLRGIRKA 266
           I  I + + A+     V+ L      G+RKA
Sbjct: 237 IDHIANADTAEQAIDGVLSLCAALADGVRKA 267


Lambda     K      H
   0.321    0.137    0.389 

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: 238
Number of extensions: 10
Number of successful extensions: 2
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: 269
Length of database: 270
Length adjustment: 25
Effective length of query: 244
Effective length of database: 245
Effective search space:    59780
Effective search space used:    59780
Neighboring words threshold: 11
Window for multiple hits: 40
X1: 16 ( 7.4 bits)
X2: 38 (14.6 bits)
X3: 64 (24.7 bits)
S1: 41 (21.8 bits)
S2: 47 (22.7 bits)

Align candidate Pf6N2E2_4047 (Tryptophan synthase alpha chain (EC 4.2.1.20))
to HMM TIGR00262 (trpA: tryptophan synthase, alpha subunit (EC 4.2.1.20))

# 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/TIGR00262.hmm
# target sequence database:        /tmp/gapView.1788.genome.faa
# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -

Query:       TIGR00262  [M=256]
Accession:   TIGR00262
Description: trpA: tryptophan synthase, alpha subunit
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
    ------- ------ -----    ------- ------ -----   ---- --  --------                                      -----------
    5.5e-82  260.5   0.0    6.1e-82  260.3   0.0    1.0  1  lcl|FitnessBrowser__pseudo6_N2E2:Pf6N2E2_4047  Tryptophan synthase alpha chain 


Domain annotation for each sequence (and alignments):
>> lcl|FitnessBrowser__pseudo6_N2E2:Pf6N2E2_4047  Tryptophan synthase alpha chain (EC 4.2.1.20)
   #    score  bias  c-Evalue  i-Evalue hmmfrom  hmm to    alifrom  ali to    envfrom  env to     acc
 ---   ------ ----- --------- --------- ------- -------    ------- -------    ------- -------    ----
   1 !  260.3   0.0   6.1e-82   6.1e-82       1     254 [.       8     259 ..       8     261 .. 0.98

  Alignments for each domain:
  == domain 1  score: 260.3 bits;  conditional E-value: 6.1e-82
                                      TIGR00262   1 fetlkkkeekafvpFvtagdPdleksleiiktlvkaGadalElGvpfsDPlaDGptiqaaelRA 64 
                                                    f++lk+++  a+v+FvtagdPd+++sl i+k l  aGad++ElG+pf DP+aDGp+iq a+ RA
  lcl|FitnessBrowser__pseudo6_N2E2:Pf6N2E2_4047   8 FAELKEQNRAALVTFVTAGDPDYDTSLAILKGLPAAGADVIELGMPFTDPMADGPAIQLANIRA 71 
                                                    7899************************************************************ PP

                                      TIGR00262  65 lkagvkvekalellkkvrekasniPivlltyynlifkkgveeFyakakeagvdgvlvaDlPlee 128
                                                    l a  +++k+l++++++r+++s+ P+vl+ y+n+i+++gv+ F+ +a e+gvdg++v+D+P e 
  lcl|FitnessBrowser__pseudo6_N2E2:Pf6N2E2_4047  72 LGAQQNLAKTLQMVREFRKDNSETPLVLMGYFNPIHRYGVQRFIGEALESGVDGLIVVDMPPEH 135
                                                    **************************************************************** PP

                                      TIGR00262 129 addlleaakkhgvkqiflvaPtaeeerlkkiaekseGfvYlvsvaGvtgarerveeevkelikk 192
                                                    +++l e a+  g++ i l +Pt+++ rl ++ + s+GfvY vsvaGvtga     e+v+e++++
  lcl|FitnessBrowser__pseudo6_N2E2:Pf6N2E2_4047 136 NSELCEPAQAAGLDFIRLTTPTTDDVRLPTVLNGSSGFVYYVSVAGVTGAGAATLEHVEEAVAR 199
                                                    **************************************************************** PP

                                      TIGR00262 193 vkalskkPvlvGFGiskkeqvkelkelgadgvivGsAlvkiieeklddeekaleeleefvke 254
                                                    ++++++ P+ +GFGi  +eq+ ++  l adgv+vGsAl++ i++  d+ e+a++ + +++ +
  lcl|FitnessBrowser__pseudo6_N2E2:Pf6N2E2_4047 200 LRRHTDLPISIGFGIRTPEQAAAIARL-ADGVVVGSALIDHIANA-DTAEQAIDGVLSLCAA 259
                                                    *************************99.9*************998.9999999887777665 PP



Internal pipeline statistics summary:
-------------------------------------
Query model(s):                            1  (256 nodes)
Target sequences:                          1  (270 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: 0.01u 0.01s 00:00:00.02 Elapsed: 00:00:00.00
# Mc/sec: 7.16
//
[ok]

This GapMind analysis is from Apr 09 2024. The underlying query database was built on Apr 09 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