ABCD1 p.Arg17Trp
| Predicted by SNAP2: | A: N (61%), C: N (57%), D: D (71%), E: D (53%), F: D (75%), G: D (53%), H: N (61%), I: N (57%), K: N (97%), L: N (57%), M: N (61%), N: N (57%), P: D (59%), Q: N (82%), S: N (66%), T: N (66%), V: N (61%), W: D (80%), Y: D (71%), |
| Predicted by PROVEAN: | A: N, C: D, D: D, E: N, F: D, G: N, H: N, I: D, K: N, L: D, M: D, N: N, P: N, Q: N, S: N, T: N, V: D, W: D, Y: D, |
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[hide] Purification and characterization of cytosolic fru... Arch Biochem Biophys. 1998 Jul 15;355(2):189-96. Hodgson RJ, Plaxton WC
Purification and characterization of cytosolic fructose-1, 6-bisphosphate aldolase from endosperm of germinated castor oil seeds.
Arch Biochem Biophys. 1998 Jul 15;355(2):189-96., [PMID:9675026]
Abstract [show]
Cytosolic fructose-1,6-bisphosphate (FBP) aldolase (ALDc) from the endosperm of 4- to 5-day germinated castor oil seeds (COS) has been purified 83-fold to electrophoretic homogeneity and a final specific activity of 2.5 micromol FBP cleaved/min/mg protein. SDS-PAGE and denaturing isoelectric focusing of the final preparation revealed a single protein-staining band of 40 kDa and pI value 7.2. The native Mr was determined by gel-filtration chromatography and multiangle laser light scattering to be 160-175 kDa, indicating that the enzyme is homotetrameric. The enzyme (a) is a class I aldolase, since EDTA or Mg2+ had no effect on its activity; and (b) was relatively heat stable and had an activation energy of 100 kJ/mol. It exhibited a broad pH-activity optima of 7.2, a relatively high affinity for FBP (Km = 0.16 microM), and a forward:reverse Vmax ratio of 0.77. Rabbit anti-(COS ALDc) antibodies cross-reacted with COS ALDc, but not with the corresponding plastidic isozyme. Time-course studies revealed that (a) the increase in total ALD activity that occurs during COS development and early germination coincides with an increase in ALDc concentration and (b) the latter stages of COS maturation and germination are accompanied by marked reductions in ALD activity and ALDc concentration. The most significant elevation in ALDc concentration occurred over the first 4 days of germination when COS initiates the gluconeogenic conversion of P-enolpyruvate and triose-P, derived from reserve triacylglycerols, into the sucrose required to support early seedling growth.
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166 Immunoremoval was performed with 0.008 unit of each ALD as described under Materials and Methods. TABLE II Amino Acid Composition of Germinated Castor Seed Cytosolic Aldolase Amino acid Residues per 40-kDa subunit Asx 35 Thr 23 Ser 14 Glx 46 Pro 20 Gly 34 Ala 46 Cys 7 Val 24 Met 2 Ile 14 Leu 39 Tyr 12 Phe 6 His 5 Lys 22 Arg 17 Trp 1 (4, 6, 8, 9, 11) the purified COS ALDc showed a very broad pH/activity profile for the forward reaction, with a maximum occurring at about pH 7.2 and near-maximal activity occurring in the pH range 6.4 to 8.0.
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ABCD1 p.Arg17Trp 9675026:166:322
status: NEW165 Immunoremoval was performed with 0.008 unit of each ALD as described under Materials and Methods. TABLE II Amino Acid Composition of Germinated Castor Seed Cytosolic Aldolase Amino acid Residues per 40-kDa subunit Asx 35 Thr 23 Ser 14 Glx 46 Pro 20 Gly 34 Ala 46 Cys 7 Val 24 Met 2 Ile 14 Leu 39 Tyr 12 Phe 6 His 5 Lys 22 Arg 17 Trp 1 (4, 6, 8, 9, 11) the purified COS ALDc showed a very broad pH/activity profile for the forward reaction, with a maximum occurring at about pH 7.2 and near-maximal activity occurring in the pH range 6.4 to 8.0.
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ABCD1 p.Arg17Trp 9675026:165:322
status: NEW