Alignments for a candidate for PAH in Caulobacter crescentus NA1000

GapMind for catabolism of small carbon sources

Alignments for a candidate for PAH in Caulobacter crescentus NA1000

Align phenylalanine 4-monooxygenase (EC 1.14.16.1) (characterized)
to candidate CCNA_01684 CCNA_01684 phenylalanine-4-hydroxylase

Query= BRENDA::P30967
         (297 letters)



>FitnessBrowser__Caulo:CCNA_01684
          Length = 294

 Score =  339 bits (870), Expect = 4e-98
 Identities = 154/261 (59%), Positives = 199/261 (76%)

Query: 26  DFTLPQPLDRYSAEDHATWATLYQRQCKLLPGRACDEFMEGLERLEVDADRVPDFNKLNQ 85
           D+T+ Q  + Y+  +H  W TLY+RQ  +L GRACDEFM GL+ L++    +PDF ++N+
Sbjct: 17  DWTIDQGWETYTQAEHDVWITLYERQTDMLHGRACDEFMRGLDALDLHRSGIPDFARINE 76

Query: 86  KLMAATGWKIVAVPGLIPDDVFFEHLANRRFPVTWWLREPHQLDYLQEPDVFHDLFGHVP 145
           +L   TGW +VAVPGL+PDDVFF+HLANRRFP   ++R+PH+LDYLQEPD+FHD+FGHVP
Sbjct: 77  ELKRLTGWTVVAVPGLVPDDVFFDHLANRRFPAGQFIRKPHELDYLQEPDIFHDVFGHVP 136

Query: 146 LLINPVFADYLEAYGKGGVKAKALGALPMLARLYWYTVEFGLINTPAGMRIYGAGILSSK 205
           +L +PVFADY++AYG+GG +A  LG L  LARLYWYTVEFGL+NTPAG+RIYGAGI+SS+
Sbjct: 137 MLTDPVFADYMQAYGEGGRRALGLGRLANLARLYWYTVEFGLMNTPAGLRIYGAGIVSSR 196

Query: 206 SESIYCLDSASPNRVGFDLMRIMNTRYRIDTFQKTYFVIDSFKQLFDATAPDFAPLYLQL 265
           +ESI+ LD  SPNR+GFDL R+M T YRID FQ+ YFVIDS + L + T  DF  +Y +L
Sbjct: 197 TESIFALDDPSPNRIGFDLERVMRTLYRIDDFQQVYFVIDSIQTLQEVTLRDFGAIYERL 256

Query: 266 ADAQPWGAGDVAPDDLVLNAG 286
           A     G  ++ P D VL  G
Sbjct: 257 ASVSDIGVAEIVPGDAVLTRG 277


Lambda     K      H
   0.323    0.140    0.445 

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: 327
Number of extensions: 7
Number of successful extensions: 1
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: 297
Length of database: 294
Length adjustment: 26
Effective length of query: 271
Effective length of database: 268
Effective search space:    72628
Effective search space used:    72628
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 (21.9 bits)
S2: 48 (23.1 bits)

Align candidate CCNA_01684 CCNA_01684 (phenylalanine-4-hydroxylase)
to HMM TIGR01267 (phhA: phenylalanine-4-hydroxylase (EC 1.14.16.1))

# 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.carbon/TIGR01267.hmm
# target sequence database:        /tmp/gapView.5704.genome.faa
# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -

Query:       TIGR01267  [M=248]
Accession:   TIGR01267
Description: Phe4hydrox_mono: phenylalanine-4-hydroxylase
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
    ------- ------ -----    ------- ------ -----   ---- --  --------                             -----------
   1.1e-110  354.7   0.0   1.3e-110  354.5   0.0    1.0  1  lcl|FitnessBrowser__Caulo:CCNA_01684  CCNA_01684 phenylalanine-4-hydro


Domain annotation for each sequence (and alignments):
>> lcl|FitnessBrowser__Caulo:CCNA_01684  CCNA_01684 phenylalanine-4-hydroxylase
   #    score  bias  c-Evalue  i-Evalue hmmfrom  hmm to    alifrom  ali to    envfrom  env to     acc
 ---   ------ ----- --------- --------- ------- -------    ------- -------    ------- -------    ----
   1 !  354.5   0.0  1.3e-110  1.3e-110       1     242 [.      17     259 ..      17     265 .. 0.98

  Alignments for each domain:
  == domain 1  score: 354.5 bits;  conditional E-value: 1.3e-110
                             TIGR01267   1 dftvaqdldryseeehavwatlidrqlkllegracdeyldGveklgldadripdleevneklraltGwkivav 73 
                                           d+t++q+++ y++ eh vw tl++rq+ +l+gracde++ G++ l+l+   ipd+  +ne l++ltGw +vav
  lcl|FitnessBrowser__Caulo:CCNA_01684  17 DWTIDQGWETYTQAEHDVWITLYERQTDMLHGRACDEFMRGLDALDLHRSGIPDFARINEELKRLTGWTVVAV 89 
                                           6799********************************************************************* PP

                             TIGR01267  74 pglipadvffehlanrrfpvttflrtpeeldylqepdvfhdlfGhvpllsnpvfadfleayGkkgvkakalga 146
                                           pgl+p+dvff+hlanrrfp  +f+r+p+eldylqepd+fhd+fGhvp+l++pvfad+++ayG +g +a  lg+
  lcl|FitnessBrowser__Caulo:CCNA_01684  90 PGLVPDDVFFDHLANRRFPAGQFIRKPHELDYLQEPDIFHDVFGHVPMLTDPVFADYMQAYGEGGRRALGLGR 162
                                           ************************************************************************9 PP

                             TIGR01267 147 a.llarlywytvefGlvetaaglriyGaGilsskkelvyaleskeplrvafdllevmrtryridklqkayfvl 218
                                              larlywytvefGl++t+aglriyGaGi+ss +e+++al+  +p+r++fdl++vmrt yrid +q++yfv+
  lcl|FitnessBrowser__Caulo:CCNA_01684 163 LaNLARLYWYTVEFGLMNTPAGLRIYGAGIVSSRTESIFALDDPSPNRIGFDLERVMRTLYRIDDFQQVYFVI 235
                                           879********************************************************************** PP

                             TIGR01267 219 pslkrlfdaaqedfealvaeakdl 242
                                           +s++ l +++  df a++ +++  
  lcl|FitnessBrowser__Caulo:CCNA_01684 236 DSIQTLQEVTLRDFGAIYERLASV 259
                                           ******************998765 PP



Internal pipeline statistics summary:
-------------------------------------
Query model(s):                            1  (248 nodes)
Target sequences:                          1  (294 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: 9.48
//
[ok]

This GapMind analysis is from Sep 17 2021. The underlying query database was built on Sep 17 2021.

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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

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 catabolism of small carbon sources