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Indian Journal of Biochemistry and Biophysics (IJBB) >
IJBB Vol.40 [2003] >
IJBB Vol.40(2) [April 2003] >
| Title: | Effect of gastrointestinal proteases on purified human intrinsic factor-vitamin B12 (IF-B12) complex |
| Authors: | Srikumar, K
Premalatha, R |
| Issue Date: | Apr-2003 |
| Publisher: | CSIR |
| Abstract: | Intrinsic factor (IF) from human gastric juice was purified and complexed with vitamin B12 (IF-B12 complex) on Sepharose-vitamin B12 affinity matrix. By labeling studies, using [57Co] vitamin B12 and 125I, the specific B12 binding activity of IF was found to be 23 μgB12/mg protein, and the molecular size by gel filtration 60 kDa. Proteolysis of the IF-B12 complex by sequential treatment with pepsin, trypsin, α-chymotrypsin and carboxypeptidase A, followed by chromatography of proteolysed complex and IF-B12 showed higher mobility of proteolysed fraction. Gel filtration, however, showed same molecular size for both proteolysed and the IF-B12 complex. On SDS-PAGE, purified IF-B12 appeared as a single band of 60 kDa. The proteolysed complex had higher mobility on SDS-PAGE and did not bind to zirconium phosphate gel. Immunodiffusion with rabbit antisera had positive reaction with IF-B12, but there was no reaction with the proteolysed sample. |
| Page(s): | 139-142 |
| ISSN: | 0301-1208 |
| Source: | IJBB Vol.40(2) [April 2003]
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