Isolation of a crystal matrix protein associated with calcium oxalate precipitation in vacuoles of specialized cells.
The formation of calcium (Ca) oxalate crystals is considered to be a high-capacity mechanism for regulating Ca in many plants. Ca oxalate precipitation is not a stochastic process, suggesting the involvement of specific biochemical and cellular mechanisms. Microautoradiography of water lettuce (Pistia stratiotes) tissue exposed to 3H-glutamate showed incorporation into developing crystals, indicating potential acidic proteins associated with the crystals. Dissolution of crystals leaves behind a crystal-shaped matrix "ghost" that is capable of precipitation of Ca oxalate in the original crystal morphology. To assess whether this matrix has a protein component, purified crystals were isolated and analyzed for internal protein. Polyacrylamide gel electrophoresis revealed the presence of one major polypeptide of about 55 kD and two minor species of 60 and 63 kD. Amino acid analysis indicates the matrix protein is relatively high in acidic amino acids, a feature consistent with its solubility in formic acid but not at neutral pH. 45Ca-binding assays demonstrated the matrix protein has a strong affinity for Ca. Immunocytochemical localization using antibody raised to the isolated protein showed that the matrix protein is specific to crystal-forming cells. Within the vacuole, the surface and internal structures of two morphologically distinct Ca oxalate crystals, raphide and druse, were labeled by the antimatrix protein serum, as were the surfaces of isolated crystals. These results demonstrate that a specific Ca-binding protein exists as an integral component of Ca oxalate crystals, which holds important implications with respect to regulation of crystal formation.
Li
Xingxiang
X
Department of Genetics and Cell Biology, Washington State University, Pullman, WA 99164-4236, USA.
Zhang
Dianzhong
D
Lynch-Holm
Valerie J
VJ
Okita
Thomas W
TW
Franceschi
Vincent R
VR
D000596:Amino Acids / Q000032:analysis; D029064:Araceae / Q000378:metabolism; D002118:Calcium / Q000502:physiology; D002129:Calcium Oxalate / Q000378:metabolism*; D003460:Crystallization; D010940:Plant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D014617:Vacuoles / Q000378:metabolism*
D000596:Amino Acids; D010940:Plant Proteins; D011506:Proteins; D002129:Calcium Oxalate; D002118:Calcium