TY - JOUR
T1 - Biochemical characterization of Acacia schweinfurthii serine proteinase inhibitor
AU - Odei-Addo, Frank
AU - Frost, Carminita
AU - Smith, Nanette
AU - Ogawa, Tomohisa
AU - Muramoto, Koji
AU - Oliva, Maria Luiza Vilela
AU - Gráf, László
AU - Naude, Ryno
N1 - Funding Information:
This work was supported by the National Research Foundation (South Africa), the Hungarian Science Foundation (Hungary), the FAPESP (09/53766-9) (Brazil) and the Nelson Mandela Metropolitan University.
Publisher Copyright:
© 2014 Informa UK Ltd.
PY - 2014/10/1
Y1 - 2014/10/1
N2 - One of the many control mechanisms of serine proteinases is their specific inhibition by protein proteinase inhibitors. An extract of Acacia schweinfurthii was screened for potential serine proteinase inhibition. It was successfully purified to homogeneity by precipitating with 80% (v/v) acetone and sequential chromatographic steps, including ion-exchange, affinity purification and reversed-phase high performance liquid chromatography. Reducing sodium dodecyl sulphate polyacrylamide gel electrophoresis conditions revealed an inhibitor (ASTI) consisting of two polypeptide chains A and B of approximate molecular weights of 16 and 10kDa, respectively, and under non-reducing conditions, 26kDa was observed. The inhibitor was shown to inhibit bovine trypsin (Ki of 3.45nM) at an approximate molar ratio of inhibitor:trypsin (1:1). The A- and B-chains revealed complete sequences of 140 and 40 amino acid residues, respectively. Sequence similarity (70%) was reported between ASTI A-chain and ACTI A-chain (Acacia confusa) using ClustalW. The B-chain produced a 76% sequence similarity between ASTI and Leucaena leucocephala trypsin inhibitor.
AB - One of the many control mechanisms of serine proteinases is their specific inhibition by protein proteinase inhibitors. An extract of Acacia schweinfurthii was screened for potential serine proteinase inhibition. It was successfully purified to homogeneity by precipitating with 80% (v/v) acetone and sequential chromatographic steps, including ion-exchange, affinity purification and reversed-phase high performance liquid chromatography. Reducing sodium dodecyl sulphate polyacrylamide gel electrophoresis conditions revealed an inhibitor (ASTI) consisting of two polypeptide chains A and B of approximate molecular weights of 16 and 10kDa, respectively, and under non-reducing conditions, 26kDa was observed. The inhibitor was shown to inhibit bovine trypsin (Ki of 3.45nM) at an approximate molar ratio of inhibitor:trypsin (1:1). The A- and B-chains revealed complete sequences of 140 and 40 amino acid residues, respectively. Sequence similarity (70%) was reported between ASTI A-chain and ACTI A-chain (Acacia confusa) using ClustalW. The B-chain produced a 76% sequence similarity between ASTI and Leucaena leucocephala trypsin inhibitor.
KW - Acacia schweinfurthii
KW - Amino acid sequence
KW - Proteinase inhibitors
KW - Trypsin inhibitor
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U2 - 10.3109/14756366.2013.836642
DO - 10.3109/14756366.2013.836642
M3 - Article
C2 - 24090421
AN - SCOPUS:84907055983
SN - 1475-6366
VL - 29
SP - 633
EP - 638
JO - Journal of Enzyme Inhibition and Medicinal Chemistry
JF - Journal of Enzyme Inhibition and Medicinal Chemistry
IS - 5
ER -