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P00693

- AMY1_HORVU

UniProt

P00693 - AMY1_HORVU

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Protein

Alpha-amylase type A isozyme

Gene

AMY1.1

Organism
Hordeum vulgare (Barley)
Status
Reviewed - Annotation score: 5 out of 5- Experimental evidence at protein leveli

Functioni

Catalytic activityi

Endohydrolysis of (1->4)-alpha-D-glucosidic linkages in polysaccharides containing three or more (1->4)-alpha-linked D-glucose units.3 Publications

Cofactori

Binds 3 calcium ions per subunit.

Sites

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Metal bindingi116 – 1161Calcium 1
Metal bindingi133 – 1331Calcium 2
Metal bindingi136 – 1361Calcium 2; via carbonyl oxygen
Metal bindingi138 – 1381Calcium 2; via carbonyl oxygen
Metal bindingi142 – 1421Calcium 2
Metal bindingi152 – 1521Calcium 3
Metal bindingi163 – 1631Calcium 1
Metal bindingi166 – 1661Calcium 1; via carbonyl oxygen
Metal bindingi167 – 1671Calcium 3
Metal bindingi168 – 1681Calcium 3; via carbonyl oxygen
Metal bindingi171 – 1711Calcium 3; via carbonyl oxygen
Metal bindingi173 – 1731Calcium 1
Metal bindingi173 – 1731Calcium 3
Active sitei204 – 2041Nucleophile1 Publication
Metal bindingi208 – 2081Calcium 1; via carbonyl oxygen
Active sitei229 – 2291Proton donor1 Publication
Binding sitei231 – 2311Substrate
Binding sitei233 – 2331Substrate
Binding sitei251 – 2511Substrate
Binding sitei258 – 2581Substrate
Binding sitei295 – 2951Substrate
Binding sitei314 – 3141Substrate
Sitei315 – 3151Transition state stabilizer
Binding sitei320 – 3201Substrate
Binding sitei399 – 3991Substrate
Binding sitei426 – 4261Substrate

GO - Molecular functioni

  1. alpha-amylase activity Source: UniProtKB-EC
  2. calcium ion binding Source: InterPro

GO - Biological processi

  1. carbohydrate metabolic process Source: UniProtKB-KW
Complete GO annotation...

Keywords - Molecular functioni

Glycosidase, Hydrolase

Keywords - Biological processi

Carbohydrate metabolism, Germination

Keywords - Ligandi

Calcium, Metal-binding

Enzyme and pathway databases

BRENDAi3.2.1.1. 2687.

Protein family/group databases

CAZyiGH13. Glycoside Hydrolase Family 13.

Names & Taxonomyi

Protein namesi
Recommended name:
Alpha-amylase type A isozyme (EC:3.2.1.1)
Alternative name(s):
1,4-alpha-D-glucan glucanohydrolase
AMY1
Low pI alpha-amylase
Gene namesi
Name:AMY1.1
OrganismiHordeum vulgare (Barley)
Taxonomic identifieri4513 [NCBI]
Taxonomic lineageiEukaryotaViridiplantaeStreptophytaEmbryophytaTracheophytaSpermatophytaMagnoliophytaLiliopsidaPoalesPoaceaeBEP cladePooideaeTriticeaeHordeum

Organism-specific databases

GrameneiP00693.

Subcellular locationi

GO - Cellular componenti

  1. extracellular region Source: UniProtKB-KW
Complete GO annotation...

Keywords - Cellular componenti

Secreted

Pathology & Biotechi

Mutagenesis

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Mutagenesisi117 – 1171H → N: 20-fold decrease in activity. 1 Publication
Mutagenesisi129 – 1291Y → A: Reduces affinity for starch granules 3-fold. 1 Publication
Mutagenesisi204 – 2041D → A or N: Loss of activity. 2 Publications
Mutagenesisi229 – 2291E → Q: Loss of activity. 1 Publication
Mutagenesisi302 – 3021W → A: Over 10-fold decrease in affinity for starch granules. Abolishes binding of starch granules and reduces activity towards starch granules by 99%; when associated with A-303 and A-404. 1 Publication
Mutagenesisi303 – 3031W → A: Over 10-fold decrease in affinity for starch granules. Abolishes binding of starch granules and reduces activity towards starch granules by 99%; when associated with A-302 and A-404. 2 Publications
Mutagenesisi314 – 3141H → N: 10-fold decrease in activity. 1 Publication
Mutagenesisi315 – 3151D → N: Loss of activity. 1 Publication
Mutagenesisi404 – 4041Y → A: Reduces affinity for starch granules 9-fold and reduces activity by 90%. Abolishes binding of starch granules and reduces activity towards starch granules by 99%; when associated with A-302 and A-303. 3 Publications
Mutagenesisi404 – 4041Y → M: Abolishes binding to cyclodextrin-Sepharose and strongly reduces enzyme activity. 3 Publications
Mutagenesisi419 – 4191H → A: Slightly decreased catalytic activity. 1 Publication

PTM / Processingi

Molecule processing

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Signal peptidei1 – 2424Add
BLAST
Chaini25 – 438414Alpha-amylase type A isozymePRO_0000001404Add
BLAST

Expressioni

Developmental stagei

Production of alpha-amylase is hormonally regulated. Germinating embryos produce the hormone gibberellic acid, which within 10 hours stimulates the aleurone cells covering the endosperm of the seed to produce alpha-amylase. The enzyme then degrades the starch within the endosperm for use by the developing plant embryo.

Gene expression databases

GenevestigatoriP00693.

Interactioni

Subunit structurei

Monomer.2 Publications

Structurei

Secondary structure

1
438
Legend: HelixTurnBeta strand
Show more details
Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Beta strandi28 – 303
Helixi36 – 383
Helixi43 – 486
Helixi51 – 566
Beta strandi61 – 644
Beta strandi70 – 723
Beta strandi75 – 784
Helixi84 – 863
Helixi92 – 10413
Beta strandi108 – 1136
Beta strandi121 – 1233
Beta strandi129 – 1313
Beta strandi135 – 1395
Helixi145 – 1473
Turni153 – 1553
Helixi179 – 19416
Beta strandi200 – 2034
Helixi206 – 2083
Helixi211 – 22111
Beta strandi226 – 2283
Beta strandi240 – 2423
Helixi247 – 26014
Helixi262 – 2643
Beta strandi265 – 2706
Helixi272 – 28110
Turni282 – 2843
Helixi286 – 2894
Helixi299 – 3024
Helixi304 – 3063
Beta strandi307 – 3115
Turni314 – 3163
Turni318 – 3203
Helixi327 – 3293
Helixi330 – 33910
Beta strandi340 – 3478
Helixi348 – 3525
Beta strandi353 – 3553
Helixi357 – 36913
Beta strandi378 – 3847
Beta strandi387 – 3926
Turni393 – 3953
Beta strandi396 – 4027
Helixi407 – 4093
Beta strandi415 – 4206
Beta strandi423 – 4275

3D structure databases

Select the link destinations:
PDBe
RCSB PDB
PDBj
Links Updated
EntryMethodResolution (Å)ChainPositionsPDBsum
1HT6X-ray1.50A25-429[»]
1P6WX-ray2.00A25-429[»]
1RP8X-ray2.00A25-429[»]
1RP9X-ray2.00A25-429[»]
1RPKX-ray2.00A25-429[»]
2QPSX-ray2.20A25-429[»]
2QPUX-ray1.70A/B/C25-429[»]
3BSGX-ray1.95A25-438[»]
3BSHX-ray3.00A25-438[»]
ProteinModelPortaliP00693.
SMRiP00693. Positions 25-428.
ModBaseiSearch...
MobiDBiSearch...

Miscellaneous databases

EvolutionaryTraceiP00693.

Family & Domainsi

Region

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Regioni69 – 713Substrate binding
Regioni76 – 772Substrate binding
Regioni202 – 2076Substrate binding
Regioni301 – 3033Substrate binding
Regioni404 – 4063Substrate binding
Regioni416 – 4227Substrate binding

Sequence similaritiesi

Belongs to the glycosyl hydrolase 13 family.Curated

Keywords - Domaini

Signal

Family and domain databases

Gene3Di2.60.40.1180. 1 hit.
3.20.20.80. 1 hit.
InterProiIPR012850. A-amylase_bs_C.
IPR013775. A-amylase_pln.
IPR006046. Alpha_amylase.
IPR015902. Glyco_hydro_13.
IPR013780. Glyco_hydro_13_b.
IPR006047. Glyco_hydro_13_cat_dom.
IPR006589. Glyco_hydro_13_sub_cat_dom.
IPR013781. Glyco_hydro_catalytic_dom.
IPR017853. Glycoside_hydrolase_SF.
[Graphical view]
PANTHERiPTHR10357. PTHR10357. 1 hit.
PfamiPF07821. Alpha-amyl_C2. 1 hit.
PF00128. Alpha-amylase. 1 hit.
[Graphical view]
PIRSFiPIRSF001028. Alph-amls_plant. 1 hit.
PRINTSiPR00110. ALPHAAMYLASE.
SMARTiSM00642. Aamy. 1 hit.
SM00810. Alpha-amyl_C2. 1 hit.
[Graphical view]
SUPFAMiSSF51445. SSF51445. 1 hit.

Sequencei

Sequence statusi: Complete.

Sequence processingi: The displayed sequence is further processed into a mature form.

P00693-1 [UniParc]FASTAAdd to Basket

« Hide

        10         20         30         40         50
MGKNGSLCCF SLLLLLLLAG LASGHQVLFQ GFNWESWKQS GGWYNMMMGK
60 70 80 90 100
VDDIAAAGVT HVWLPPPSHS VSNEGYMPGR LYDIDASKYG NAAELKSLIG
110 120 130 140 150
ALHGKGVQAI ADIVINHRCA DYKDSRGIYC IFEGGTSDGR LDWGPHMICR
160 170 180 190 200
DDTKYSDGTA NLDTGADFAA APDIDHLNDR VQRELKEWLL WLKSDLGFDA
210 220 230 240 250
WRLDFARGYS PEMAKVYIDG TSPSLAVAEV WDNMATGGDG KPNYDQDAHR
260 270 280 290 300
QNLVNWVDKV GGAASAGMVF DFTTKGILNA AVEGELWRLI DPQGKAPGVM
310 320 330 340 350
GWWPAKAATF VDNHDTGSTQ AMWPFPSDKV MQGYAYILTH PGIPCIFYDH
360 370 380 390 400
FFNWGFKDQI AALVAIRKRN GITATSALKI LMHEGDAYVA EIDGKVVVKI
410 420 430
GSRYDVGAVI PAGFVTSAHG NDYAVWEKNG AAATLQRS
Length:438
Mass (Da):47,796
Last modified:July 21, 1986 - v1
Checksum:i2393FDAC51E80F51
GO

Sequence databases

Select the link destinations:
EMBL
GenBank
DDBJ
Links Updated
J01236 mRNA. Translation: AAA32929.1.
PIRiA00846. ALBH.
UniGeneiHv.68.

Cross-referencesi

Sequence databases

Select the link destinations:
EMBL
GenBank
DDBJ
Links Updated
J01236 mRNA. Translation: AAA32929.1 .
PIRi A00846. ALBH.
UniGenei Hv.68.

3D structure databases

Select the link destinations:
PDBe
RCSB PDB
PDBj
Links Updated
Entry Method Resolution (Å) Chain Positions PDBsum
1HT6 X-ray 1.50 A 25-429 [» ]
1P6W X-ray 2.00 A 25-429 [» ]
1RP8 X-ray 2.00 A 25-429 [» ]
1RP9 X-ray 2.00 A 25-429 [» ]
1RPK X-ray 2.00 A 25-429 [» ]
2QPS X-ray 2.20 A 25-429 [» ]
2QPU X-ray 1.70 A/B/C 25-429 [» ]
3BSG X-ray 1.95 A 25-438 [» ]
3BSH X-ray 3.00 A 25-438 [» ]
ProteinModelPortali P00693.
SMRi P00693. Positions 25-428.
ModBasei Search...
MobiDBi Search...

Protein family/group databases

CAZyi GH13. Glycoside Hydrolase Family 13.

Protocols and materials databases

Structural Biology Knowledgebase Search...

Organism-specific databases

Gramenei P00693.

Enzyme and pathway databases

BRENDAi 3.2.1.1. 2687.

Miscellaneous databases

EvolutionaryTracei P00693.

Gene expression databases

Genevestigatori P00693.

Family and domain databases

Gene3Di 2.60.40.1180. 1 hit.
3.20.20.80. 1 hit.
InterProi IPR012850. A-amylase_bs_C.
IPR013775. A-amylase_pln.
IPR006046. Alpha_amylase.
IPR015902. Glyco_hydro_13.
IPR013780. Glyco_hydro_13_b.
IPR006047. Glyco_hydro_13_cat_dom.
IPR006589. Glyco_hydro_13_sub_cat_dom.
IPR013781. Glyco_hydro_catalytic_dom.
IPR017853. Glycoside_hydrolase_SF.
[Graphical view ]
PANTHERi PTHR10357. PTHR10357. 1 hit.
Pfami PF07821. Alpha-amyl_C2. 1 hit.
PF00128. Alpha-amylase. 1 hit.
[Graphical view ]
PIRSFi PIRSF001028. Alph-amls_plant. 1 hit.
PRINTSi PR00110. ALPHAAMYLASE.
SMARTi SM00642. Aamy. 1 hit.
SM00810. Alpha-amyl_C2. 1 hit.
[Graphical view ]
SUPFAMi SSF51445. SSF51445. 1 hit.
ProtoNeti Search...

Publicationsi

  1. "Isolation and sequence analysis of a barley alpha-amylase cDNA clone."
    Rogers J.C., Milliman C.
    J. Biol. Chem. 258:8169-8174(1983) [PubMed] [Europe PMC] [Abstract]
    Cited for: NUCLEOTIDE SEQUENCE [MRNA].
    Strain: cv. Himalaya.
    Tissue: Aleurone.
  2. "Site-directed mutagenesis of histidine 93, aspartic acid 180, glutamic acid 205, histidine 290, and aspartic acid 291 at the active site and tryptophan 279 at the raw starch binding site in barley alpha-amylase 1."
    Sogaard M., Kadziola A., Haser R., Svensson B.
    J. Biol. Chem. 268:22480-22484(1993) [PubMed] [Europe PMC] [Abstract]
    Cited for: MUTAGENESIS OF HIS-117; ASP-204; GLU-229; TRP-303; HIS-314 AND ASP-315, CATALYTIC ACTIVITY, ACTIVE SITE.
  3. "Two secondary carbohydrate binding sites on the surface of barley alpha-amylase 1 have distinct functions and display synergy in hydrolysis of starch granules."
    Nielsen M.M., Bozonnet S., Seo E.S., Motyan J.A., Andersen J.M., Dilokpimol A., Abou Hachem M., Gyemant G., Naested H., Kandra L., Sigurskjold B.W., Svensson B.
    Biochemistry 48:7686-7697(2009) [PubMed] [Europe PMC] [Abstract]
    Cited for: MUTAGENESIS OF TRP-302; TRP-303 AND TYR-404.
  4. "The structure of barley alpha-amylase isozyme 1 reveals a novel role of domain C in substrate recognition and binding: a pair of sugar tongs."
    Robert X., Haser R., Gottschalk T.E., Ratajczak F., Driguez H., Svensson B., Aghajari N.
    Structure 11:973-984(2003) [PubMed] [Europe PMC] [Abstract]
    Cited for: X-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS) OF 25-428 IN COMPLEX WITH CALCIUM IONS.
  5. Cited for: X-RAY CRYSTALLOGRAPHY (2.00 ANGSTROMS) OF 25-429 IN COMPLEXES WITH CALCIUM IONS; ACARBOSE AND MALTOHEPTAOSE, CATALYTIC ACTIVITY, MUTAGENESIS OF ASP-204.
  6. "The 'pair of sugar tongs' site on the non-catalytic domain C of barley alpha-amylase participates in substrate binding and activity."
    Bozonnet S., Jensen M.T., Nielsen M.M., Aghajari N., Jensen M.H., Kramhoeft B., Willemoes M., Tranier S., Haser R., Svensson B.
    FEBS J. 274:5055-5067(2007) [PubMed] [Europe PMC] [Abstract]
    Cited for: X-RAY CRYSTALLOGRAPHY (1.70 ANGSTROMS) OF 25-429 IN COMPLEX WITH CALCIUM IONS AND ACARBOSE, MUTAGENESIS OF TYR-404.
  7. "Multi-site substrate binding and interplay in barley alpha-amylase 1."
    Nielsen M.M., Seo E.S., Bozonnet S., Aghajari N., Robert X., Haser R., Svensson B.
    FEBS Lett. 582:2567-2571(2008) [PubMed] [Europe PMC] [Abstract]
    Cited for: X-RAY CRYSTALLOGRAPHY (1.95 ANGSTROMS) IN COMPLEXES WITH CALCIUM IONS AND GLUCOSE, CATALYTIC ACTIVITY, MUTAGENESIS OF TYR-129; TYR-404 AND HIS-419.

Entry informationi

Entry nameiAMY1_HORVU
AccessioniPrimary (citable) accession number: P00693
Entry historyi
Integrated into UniProtKB/Swiss-Prot: July 21, 1986
Last sequence update: July 21, 1986
Last modified: October 29, 2014
This is version 115 of the entry and version 1 of the sequence. [Complete history]
Entry statusiReviewed (UniProtKB/Swiss-Prot)
Annotation programPlant Protein Annotation Program

Miscellaneousi

Miscellaneous

There are at least 4 types of alpha-amylase in barley.
Mutagenesis experiments indicate that His-117 and His-314 participate in transition state stabilization but not directly in catalysis.
Binds starch not only at the active site, but also via accessory binding sites on the protein surface that are important for efficient binding to starch granules and thereby increase enzyme activity.

Keywords - Technical termi

3D-structure

Documents

  1. Glycosyl hydrolases
    Classification of glycosyl hydrolase families and list of entries
  2. PDB cross-references
    Index of Protein Data Bank (PDB) cross-references
  3. SIMILARITY comments
    Index of protein domains and families

External Data

Dasty 3