Characterisation of a homogenous plant aminoaldehyde
dehydrogenase

J 2000

Characterisation of a homogenous plant aminoaldehyde dehydrogenase

ŠEBELA, Marek, Pavel PEČ, Petr GALUSZKA a Jan HAVLIŠ

Základní údaje

Originální název

Characterisation of a homogenous plant aminoaldehyde dehydrogenase

Autoři

ŠEBELA, Marek (203 Česká republika, garant), Pavel PEČ (203 Česká republika), Petr GALUSZKA (203 Česká republika) a Jan HAVLIŠ (203 Česká republika)

Vydání

Biochimica et Biophysica Acta, Amsterdam, Elsevier Science B.V. 2000, 0005-2728

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10600 1.6 Biological sciences

Stát vydavatele

Nizozemské království

Utajení

není předmětem státního či obchodního tajemství

Impakt faktor

Impact factor: 6.346

Kód RIV

RIV/00216224:14310/00:00003346

Organizační jednotka

Přírodovědecká fakulta

Klíčová slova anglicky

aminoaldehyde dehydrogenase; 3-aminopropionaldehyde; 4-aminobutyraldehyde; pea; amine oxidase

Štítky

3-aminopropionaldehyde, 4-aminobutyraldehyde, amine oxidase, aminoaldehyde dehydrogenase, pea

Změněno: 18. 1. 2007 15:20, doc. Mgr. Jan Havliš, Dr.

Anotace

V originále

According to our knowledge, this is the first purification method developed, enabling isolation of a homogeneous aminoaldehyde dehydrogenase (AMADH) from etiolated pea seedlings. The procedure involved initial purification with precipitants followed by three low pressure chromatographic steps. Partially purified enzyme was further subjected to fast protein liquid chromatography on a Mono Q column and to affinity^interaction chromatography on 5P-AMP Sepharose. Purity of the final enzyme preparation was checked by sodium dodecyl sulfate^polyacrylamide gel electrophoresis and chromatofocusing. Pea AMADH exists as a tetramer of 230 kDa in the native state, a molecular mass of one subunit was determined as 57 kDa. The enzyme was found to be an acidic protein with pI 5.4. AMADH showed a broad substrate specificity utilising various aminoaldehydes (C3^C6) as substrates. The best substrate of pea AMADH was 3- aminopropionaldehyde, the enzyme also efficiently oxidised 4-aminobutyraldehyde and g-guanidinoanalogues of the aminoaldehydes. Pea AMADH was inhibited by SH reagents, several elementary aldehydes and metal-binding agents. Although AMADH did not oxidise betaine aldehyde at all, the N-terminal amino acid sequence of the enzyme shows a high degree of homology with those of plant betaine aldehyde dehydrogenases (BADHs) of spinach, sugar beet and amaranth. Several conserved amino acids were found in comparison with BADH from cod liver of known crystal structure.

Návaznosti

GA203/99/D048, projekt VaV

Název: Purifikace rostlinné aminoaldehyddehydrogenasy a její enzymologická charakterizace

Citovat

ŠEBELA, Marek, Pavel PEČ, Petr GALUSZKA a Jan HAVLIŠ. Characterisation of a homogenous plant aminoaldehyde dehydrogenase. Biochimica et Biophysica Acta. Amsterdam: Elsevier Science B.V., 2000, -, č. 1480, s. 329-669. ISSN 0005-2728.

@article{335491,
   author = {Šebela, Marek and Peč, Pavel and Galuszka, Petr and Havliš, Jan},
   article_location = {Amsterdam},
   article_number = {1480},
   keywords = {aminoaldehyde dehydrogenase; 3-aminopropionaldehyde; 4-aminobutyraldehyde; pea; amine oxidase},
   language = {eng},
   issn = {0005-2728},
   journal = {Biochimica et Biophysica Acta},
   title = {Characterisation of a homogenous plant aminoaldehyde dehydrogenase},
   volume = {-},
   year = {2000}
}

TY  - JOUR
ID  - 335491
AU  - Šebela, Marek - Peč, Pavel - Galuszka, Petr - Havliš, Jan
PY  - 2000
TI  - Characterisation of a homogenous plant aminoaldehyde dehydrogenase
JF  - Biochimica et Biophysica Acta
VL  - -
IS  - 1480
SP  - 329
EP  - 329
PB  - Elsevier Science B.V.
SN  - 00052728
KW  - aminoaldehyde dehydrogenase
KW  - 3-aminopropionaldehyde
KW  - 4-aminobutyraldehyde
KW  - pea
KW  - amine oxidase
N2  - According to our knowledge, this is the first purification method developed, enabling isolation of a homogeneous aminoaldehyde dehydrogenase (AMADH) from etiolated pea seedlings. The procedure involved initial purification with precipitants followed by three low pressure chromatographic steps. Partially purified enzyme was further subjected to fast protein liquid chromatography on a Mono Q column and to affinity^interaction chromatography on 5P-AMP Sepharose. Purity of the final enzyme preparation was checked by sodium dodecyl sulfate^polyacrylamide gel electrophoresis and chromatofocusing. Pea AMADH exists as a tetramer of 230 kDa in the native state, a molecular mass of one subunit was determined as 57 kDa. The enzyme was found to be an acidic protein with pI 5.4. AMADH showed a broad substrate specificity utilising various aminoaldehydes (C3^C6) as substrates. The best substrate of pea AMADH was 3- aminopropionaldehyde, the enzyme also efficiently oxidised 4-aminobutyraldehyde and g-guanidinoanalogues of the aminoaldehydes. Pea AMADH was inhibited by SH reagents, several elementary aldehydes and metal-binding agents. Although AMADH did not oxidise betaine aldehyde at all, the N-terminal amino acid sequence of the enzyme shows a high degree of homology with those of plant betaine aldehyde dehydrogenases (BADHs) of spinach, sugar beet and amaranth. Several conserved amino acids were found in comparison with BADH from cod liver of known crystal structure.
ER  -

ŠEBELA, Marek, Pavel PEČ, Petr GALUSZKA a Jan HAVLIŠ. Characterisation of a homogenous plant aminoaldehyde dehydrogenase. \textit{Biochimica et Biophysica Acta}. Amsterdam: Elsevier Science B.V., 2000, -, č.~1480, s.~329-669. ISSN~0005-2728.

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