The anchoring filament protein kalinin is synthesized and secreted as a high molecular weight precursor.
Kalinin, a recently characterized novel protein component of anchoring filaments, has been shown to be involved in keratinocyte attachment to culture substrates and to dermis in vivo, and to exist in keratinocyte-conditioned culture medium in two heterotrimeric forms of 440 and 400 kDa (Rousselle, P., Lunstrum, G.P., Keene, D.R., and Burgeson, R.E. (1991) J. Cell Biol. 114, 567-576). This study demonstrates that kalinin is initially synthesized in a cell-associated form estimated to be 460 kDa. By second dimension reduced electrophoresis, V8 protease digestion, and immunoblot analysis, we demonstrate that the cell form contains nonidentical subunits of 200, 155, and 140 kDa. The 440-kDa medium form is derived from the cell form by extracellular processing of the 200-kDa subunit to 165 kDa, a step which also occurs in skin organ culture. The 400-kDa form is derived from the 440-kDa form by extracellular processing of the 155 kDa-subunit to 105 kDa. The cell form is secreted by keratinocytes, deposited onto culture substratum, and is the form which facilitates attachment and adhesion of growing and spreading keratinocytes. It is also the form initially synthesized in skin organ culture. Kalinin purified from tissue, which appears to facilitate epithelial-mesenchymal cohesion in vivo, is closely related to the 400-kDa medium form purified from culture.
Marinkovich
M P
MP
Shriner's Hospital for Crippled Children, Portland, Oregon.
Lunstrum
G P
GP
Burgeson
R E
RE
D000650:Amnion / Q000378:metabolism*; D000818:Animals; D001345:Autoradiography; D002294:Carcinoma, Squamous Cell; D002417:Cattle; D015815:Cell Adhesion Molecules / Q000096:biosynthesis* / Q000302:isolation & purification; D002460:Cell Line; D002478:Cells, Cultured; D003545:Cysteine / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D007231:Infant, Newborn; D015603:Keratinocytes / Q000378:metabolism*; D007700:Kinetics; D046911:Macromolecular Substances; D008297:Male; D008715:Methionine / Q000378:metabolism; D008970:Molecular Weight; D009924:Organ Culture Techniques; D010446:Peptide Fragments / Q000302:isolation & purification; D011247:Pregnancy; D011498:Protein Precursors / Q000096:biosynthesis* / Q000302:isolation & purification; D012867:Skin / Q000378:metabolism*; D013462:Sulfur Radioisotopes; D000097794:Kalinin
D015815:Cell Adhesion Molecules; D003545:Cysteine; D046911:Macromolecular Substances; D008715:Methionine; D010446:Peptide Fragments; D011498:Protein Precursors; D013462:Sulfur Radioisotopes; D000097794:Kalinin