Keratins 1 and 10 or homologues as regular constituents of inner root sheath and cuticle cells in the human hair follicle.
Morphological maturation of the inner root sheath (IRS) and cuticle of the human hair follicle reveals analogies to differentiation processes in other keratinizing epithelia. Detailed biochemical analysis of respective differentiation products, however, has been largely restricted by their low solubility. Herein we provide further evidence for the existence of K1 and K10-derivatives in IRS and hair cuticle based on protein analysis of isolated fractions and immunofluorescence in situ, substantiating our earlier data (Stark, H. J., et al. Differentiation 35, 236-248 (1987)). Extracts from both compartments showed on two-dimensional (2D)-polyacrylamide gels a group of presumptive K1 and K10-turnover products in a wide pI (basic to acidic) and Mr range (56,000-65,000), named IC-I to III and IC-IV, respectively. These components (also found in nail plate) reacted with specific antibodies (to K1 and K10) on Western blots. Weak but distinctive radiolabeling of presumptive precursor spots close to authentic K1 and K10, respectively, and their presence in lower follicle fractions (distant from infundibulum) largely precluded epidermal contamination. Two-dimensional tryptic peptide maps of excised 2D spots from the IC-I to III series revealed high homology to K1, and those from IC-IV components to K10. Immunodetection in frozen sections was improved by trypsin pretreatment and showed distinguished staining for K1 and K10 in IRS ranging from the lower bulbus region up to the "keratinizing zone" of the follicle. Above, the reaction was abruptly abolished which coincides with ultrastructural "melting" of distinct filaments in the intracellular matrix. Thus, our data suggest that differentiation in these follicular compartments (IRS and cuticle) might follow common principles of keratinization.
Stark
H J
HJ
Division of Differentiation and Carcinogenesis in vitro, German Cancer Research Center, Heidelberg.
Breitkreutz
D
D
Limat
A
A
Ryle
C M
CM
Roop
D
D
Leigh
I
I
Fusenig
N
N
D005455:Fluorescent Antibody Technique; D006197:Hair / Q000737:chemistry* / Q000166:cytology / Q000648:ultrastructure; D006801:Humans; D015151:Immunoblotting; D007633:Keratins / Q000145:classification / Q000302:isolation & purification*; D010449:Peptide Mapping; D013462:Sulfur Radioisotopes; D014357:Trypsin
D013462:Sulfur Radioisotopes; D007633:Keratins; D014357:Trypsin