The characterization of recombinant mouse glandular kallikreins from E. coli.
A system has been developed for the expression in E. coli of 12 of the 14 expressed mouse submandibular gland kallikreins as cassettes subcloned directly from cDNA. Using the epidermal growth factor binding protein (mGK-9) and the gamma-subunit of nerve growth factor (mGK-3), as test cases, mature processed forms, obtained as functionally active proteins, as well as various precursor forms, were isolated. The expression system described allows rapid isolation of kallikrein protein from corresponding cDNA with yields of approximately 1.0 mg of purified protein from 10 g of initial cell paste. This expression system will facilitate structure/function studies of the mouse glandular kallikrein gene family and help elucidate the regions of the mature proteins responsible for the diverse catalytic behavior and growth factor interactions observed in this family of proteins.
Department of Biological Chemistry, College of Medicine, University of California, Irvine 92717.
M
M
Blaber
P J
PJ
Isackson
R A
RA
Bradshaw
D000595:Amino Acid Sequence; D000818:Animals; D004247:DNA / Q000302:isolation & purification; D010450:Endopeptidases / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D007610:Kallikreins / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D005810:Multigene Family; D009414:Nerve Growth Factors / Q000235:genetics*; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000235:genetics; D012995:Solubility; D013363:Submandibular Gland / Q000032:analysis*; D020840:Tissue Kallikreins
D009414:Nerve Growth Factors; D011994:Recombinant Proteins; D004247:DNA; D010450:Endopeptidases; D007610:Kallikreins; C489120:Klk13 protein, mouse; D020840:Tissue Kallikreins; C489121:Klk1b22 protein, mouse; C489119:Klk9 protein, mouse; C053223:epidermal growth factor binding protein, mouse