Presence in Human Epidermal Cells of a Soluble Protein Precursor of the CrossLinked Envelope Activation of the CrossLinking by Calcium Ions Robert H Rice and Howard Green Department of Biology Ma ID: 953378
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Cell, Vol. 18. 681-694, November 1979, Copyright 0 by MIT Presence in Human Epidermal Cells of a Soluble Protein Precursor of the Cr
oss-Linked Envelope: Activation of the Cross-Linking by Calcium Ions Robert H. Rice and Howard Green Department of Biology Massachus
etts Institute of Technology Cambridge, Massachusetts 02139 Summary Late in the terminal differentiation of epidermis and cultured ep
idermal cells, a protein envelope located beneath the plasma membrane becomes cross- linked by cellular transglutaminase. The Cl?ll
682 Trlton concentration C%) Triton concentmtion (%) Figure 1. Envelope Formation as a Function of Triton X-l 00 Concen- tration Conf
luent surface cultures (a) and trypsinized cell suspensions (b) were exposed to the indicated concentration of Triton for 4 hr and th
en extracted with 1% Time ;hours 1 4 Figure 3. Retention of Envelope Precursor in Detergent-Treated Cells Cell suspensions were tre
ated with 0.01% Triton X-l 00. (0) Control, no other additions. (0) EGTA added to 10 mM; afler 2 hr (arrow), CaCI, 60 Table 1. Puri
fication of Envelope Precursor Protein Fraction (8.0 ml) Figure 8. Fractionation of Crude Extract of Cultivated Epidermal Cells on Bi
ogel A-l 5 M The location of the envelope precursor protein was determined by precipitin formation using a specific antiserum. The pr
ecursor protein (0) eluted near the void volume (Fraction 9). Most cell protein, including lactate dehydrogenase. eluted much later.
In other - l NaCI (M) Fraction (7.5 ml) Figure IO. Fractionation of Crude Extract of Cultivated Epidermal Cells on DEAE Cellulose A
crude cell extract was applied to the column together with a marker of previously purified precursor protein labeled with ‘%-2-
amino- ethanol. The labeled precursor (0) eluted sharply at about 0.15 M NaCI. Bracket shows 10 pg and 100 pg depended on on a 10 p