Subsite mapping of human salivary α-amylase and the mutant Y151M

TY - JOUR

T1 - Subsite mapping of human salivary α-amylase and the mutant Y151M

AU - Kandra, Lili

AU - Gyémánt, Gyöngyi

AU - Remenyik, Judit

AU - Ragunath, Chandran

AU - Ramasubbu, Narayanan

N1 - Funding Information: This work was supported by OTKA T032005, and a scholarship from the Hungarian Academy of Sciences (J.R.) and the USPHS Grant DE12585 (N.R.).

PY - 2003/6/5

Y1 - 2003/6/5

N2 - This study characterizes the substrate-binding sites of human salivary α-amylase (HSA) and its Y151M mutant. It describes the first subsite maps, namely, the number of subsites, the position of cleavage sites and apparent subsite energies. The product pattern and cleavage frequencies were determined by high-performance liquid chromatography, utilizing a homologous series of chromophore-substituted maltooligosaccharides of degree of polymerization 3-10 as model substrates. The binding region of HSA is composed of four glycone and three aglycone-binding sites, while that of Tyr151Met is composed of four glycone and two aglycone-binding sites. The subsite maps show that Y151M has strikingly decreased binding energy at subsite (+2), where the mutation has occurred (-2.6 kJ/mol), compared to the binding energy at subsite (+2) of HSA (-12.0 kJ/mol).

AB - This study characterizes the substrate-binding sites of human salivary α-amylase (HSA) and its Y151M mutant. It describes the first subsite maps, namely, the number of subsites, the position of cleavage sites and apparent subsite energies. The product pattern and cleavage frequencies were determined by high-performance liquid chromatography, utilizing a homologous series of chromophore-substituted maltooligosaccharides of degree of polymerization 3-10 as model substrates. The binding region of HSA is composed of four glycone and three aglycone-binding sites, while that of Tyr151Met is composed of four glycone and two aglycone-binding sites. The subsite maps show that Y151M has strikingly decreased binding energy at subsite (+2), where the mutation has occurred (-2.6 kJ/mol), compared to the binding energy at subsite (+2) of HSA (-12.0 kJ/mol).

KW - Action pattern

KW - Binding energy

KW - Human salivary α-amylase

KW - Maltooligosaccharide

KW - Subsite map

KW - Y151M mutant

UR - https://www.scopus.com/pages/publications/0038694855

UR - https://www.scopus.com/pages/publications/0038694855#tab=citedBy

U2 - 10.1016/S0014-5793(03)00495-2

DO - 10.1016/S0014-5793(03)00495-2

M3 - Article

C2 - 12782315

SN - 0014-5793

VL - 544

SP - 194

EP - 198

JO - FEBS Letters

JF - FEBS Letters

IS - 1-3

ER -