TY - JOUR
T1 - Biochemical studies on lysin, a cell wall degrading enzyme released during fertilization in Chlamydomonas
AU - Buchanan, Marty J.
AU - Snell, William J.
N1 - Funding Information:
We thank Drs. Fred Grinnell, George Bloom, and Woodring Wright for their helpful discussions. We also thank Dr. George Bloom for his advice with the velocity sedimentation determinations. This work was supported by NIH Grant GM-25661 and NSF Grant DCB-8519845. The HPLC system was purchased with funds from an NSF Bio-instrumentation grant. M.J.B was the recipient of a grant from the Nobel Foundation.
PY - 1988/11
Y1 - 1988/11
N2 - New methods have been developed for the purification and characterization of the cell wall degrading enzyme, lysin, which is released into the medium during the mating reaction of the biflagellated alga Chlamydomonas reinhardtii. A quantitative spectrophotometric assay that detects the number of cells losing walls was used to devise a procedure for the 60-fold purification of lysin from the medium of mating gametes with a 30% yield of activity. Molecular sieve and ion exchange chromatography in combination with SDS-PAGE showed that lysin was a single polypeptide with an Mr, of 60,000. High-performance liquid chromatography and sucrose density gradient centrifugation of lysin activity were used to obtain an estimate of 66,000 D for the nondenatured molecular weight of lysin, indicating that lysin behaves as a monomer.
AB - New methods have been developed for the purification and characterization of the cell wall degrading enzyme, lysin, which is released into the medium during the mating reaction of the biflagellated alga Chlamydomonas reinhardtii. A quantitative spectrophotometric assay that detects the number of cells losing walls was used to devise a procedure for the 60-fold purification of lysin from the medium of mating gametes with a 30% yield of activity. Molecular sieve and ion exchange chromatography in combination with SDS-PAGE showed that lysin was a single polypeptide with an Mr, of 60,000. High-performance liquid chromatography and sucrose density gradient centrifugation of lysin activity were used to obtain an estimate of 66,000 D for the nondenatured molecular weight of lysin, indicating that lysin behaves as a monomer.
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U2 - 10.1016/0014-4827(88)90357-6
DO - 10.1016/0014-4827(88)90357-6
M3 - Article
C2 - 3169140
AN - SCOPUS:0023779507
SN - 0014-4827
VL - 179
SP - 181
EP - 193
JO - Experimental Cell Research
JF - Experimental Cell Research
IS - 1
ER -