Structure-function relationships of porcine pyridoxal kinase

Yun Chung Leung; H.Y. Wong; J.E. Churchich; Chun Lap Samuel Lo; F. Kwok

doi:10.1007/978-3-0348-8397-9_45

Structure-function relationships of porcine pyridoxal kinase

Yun Chung Leung, H.Y. Wong, J.E. Churchich, Chun Lap Samuel Lo, F. Kwok

Research output: Chapter in book / Conference proceeding › Chapter in an edited book (as author) › Academic research

Abstract

Pyridoxal kinase (PK) catalyzes the formation of pyridoxal-5-phosphate (PLP) from pyridoxal (PL), ATP and a divalent cation (Zn2+). So far, there is no three-dimensional structure of PK available. Site-directed mutagenesis was carried out to study the importance of three conserved residues: Tyr137, Gly242 and G1y244. The mutants (Y137F, G242A and G244A) were constructed, expressed, purified and analyzed. The results demonstrated that the mutants had much less activity but with no dramatic variation in protein stability. Tyrl 37 residue was shown to be involved in PL binding but not ATP binding. For the G244A mutant, the absence of enzyme activity was probably due to the deficiency in PL binding rather than the lack of ATP binding. In the case of the G242A mutant, it did not bind to ATP or PL.

Original language	English
Title of host publication	Biochemistry and molecular biology of vitamin B₆ and PQQ-dependent proteins
Publisher	Birkhäuser Verlag
Pages	277-280
Number of pages	4
ISBN (Electronic)	9783034883979
ISBN (Print)	9783034895491
DOIs	https://doi.org/10.1007/978-3-0348-8397-9_45
Publication status	Published - 2000

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10.1007/978-3-0348-8397-9_45

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title = "Structure-function relationships of porcine pyridoxal kinase",

abstract = "Pyridoxal kinase (PK) catalyzes the formation of pyridoxal-5-phosphate (PLP) from pyridoxal (PL), ATP and a divalent cation (Zn2+). So far, there is no three-dimensional structure of PK available. Site-directed mutagenesis was carried out to study the importance of three conserved residues: Tyr137, Gly242 and G1y244. The mutants (Y137F, G242A and G244A) were constructed, expressed, purified and analyzed. The results demonstrated that the mutants had much less activity but with no dramatic variation in protein stability. Tyrl 37 residue was shown to be involved in PL binding but not ATP binding. For the G244A mutant, the absence of enzyme activity was probably due to the deficiency in PL binding rather than the lack of ATP binding. In the case of the G242A mutant, it did not bind to ATP or PL.",

author = "Leung, {Yun Chung} and H.Y. Wong and J.E. Churchich and Lo, {Chun Lap Samuel} and F. Kwok",

year = "2000",

doi = "10.1007/978-3-0348-8397-9_45",

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TY - CHAP

T1 - Structure-function relationships of porcine pyridoxal kinase

AU - Leung, Yun Chung

AU - Wong, H.Y.

AU - Churchich, J.E.

AU - Lo, Chun Lap Samuel

AU - Kwok, F.

PY - 2000

Y1 - 2000

N2 - Pyridoxal kinase (PK) catalyzes the formation of pyridoxal-5-phosphate (PLP) from pyridoxal (PL), ATP and a divalent cation (Zn2+). So far, there is no three-dimensional structure of PK available. Site-directed mutagenesis was carried out to study the importance of three conserved residues: Tyr137, Gly242 and G1y244. The mutants (Y137F, G242A and G244A) were constructed, expressed, purified and analyzed. The results demonstrated that the mutants had much less activity but with no dramatic variation in protein stability. Tyrl 37 residue was shown to be involved in PL binding but not ATP binding. For the G244A mutant, the absence of enzyme activity was probably due to the deficiency in PL binding rather than the lack of ATP binding. In the case of the G242A mutant, it did not bind to ATP or PL.

AB - Pyridoxal kinase (PK) catalyzes the formation of pyridoxal-5-phosphate (PLP) from pyridoxal (PL), ATP and a divalent cation (Zn2+). So far, there is no three-dimensional structure of PK available. Site-directed mutagenesis was carried out to study the importance of three conserved residues: Tyr137, Gly242 and G1y244. The mutants (Y137F, G242A and G244A) were constructed, expressed, purified and analyzed. The results demonstrated that the mutants had much less activity but with no dramatic variation in protein stability. Tyrl 37 residue was shown to be involved in PL binding but not ATP binding. For the G244A mutant, the absence of enzyme activity was probably due to the deficiency in PL binding rather than the lack of ATP binding. In the case of the G242A mutant, it did not bind to ATP or PL.

U2 - 10.1007/978-3-0348-8397-9_45

DO - 10.1007/978-3-0348-8397-9_45

M3 - Chapter in an edited book (as author)

SN - 9783034895491

SP - 277

EP - 280

BT - Biochemistry and molecular biology of vitamin B₆ and PQQ-dependent proteins

PB - Birkhäuser Verlag

ER -

Structure-function relationships of porcine pyridoxal kinase

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