Autoantibody in pemphigus serum, specific for the 59 kD keratin, selectively binds the surface of keratinocytes: evidence for an extracellular keratin domain

Journal Article · Tue Sep 01 00:00:00 EDT 1987 · J. Invest. Dermatol.; (United States)

DOI:https://doi.org/10.1111/1523-1747.ep12471451· OSTI ID:6246034

Diaz, L A; Sampaio, S A; Martins, C R; Rivitti, E A; Macca, M L; Roscoe, J T; Takahashi, Y; Labib, R S; Patel, H P; Mutasim, D F

We have identified a novel IgG antikeratin autoantibody in the serum of a Brazilian pemphigus foliaceus patient (Cascas-42). This antibody is specific for the 59 kD acidic murine keratin and its 56.5 kD human counterpart (Moll's catalogue number10), and is distinct from the pemphigus antibody system. Antikeratin autoantibodies present in the Cascas-42 serum were purified by affinity chromatography with a 59 kD murine keratin-agarose column (IAP-Cascas-42 antibodies). The specificity of the IAP-Cascas-42 antibodies was tested by indirect immunofluorescence and immunoelectron microscopy against epidermal cryosections, trypsin-dissociated keratinocytes, and epidermal cell cultures. The serum was also tested with extracts from unlabeled and surface /sup 125/I-labeled keratinocytes (Iodo-Gen method) by immunoblot analysis of one- and two-dimensional polyacrylamide gel electrophoresis. The IAP-Cascas-42 antibodies bind the intercellular spaces of murine epidermis, and the cell surfaces of viable, dissociated murine keratinocytes, as well as murine epidermal cells in culture by immunofluorescence and immunoelectron microscopy. These autoantibodies did not stain cytoplasmic keratins and did not react with parallel human epidermal substrates. The Cascas-42 serum identified the 59 kD murine acidic keratin and its 56.5 kD human counterpart in epidermal extracts by two-dimensional polyacrylamide gel electrophoresis and immunoblot analysis. In addition, surface radioiodination of viable murine keratinocytes selectively labeled the 59 kD keratin suggesting that a domain of this molecule is exposed on the cell surface. The /sup 125/I-labeled 59 kD keratin was also recognized by the Cascas-42 serum by immunoblotting and autoradiography.

• Research Organization: Johns Hopkins Univ. School of Medicine, Baltimore, MD
• OSTI ID: 6246034
• Journal Information: J. Invest. Dermatol.; (United States), Vol. 89:3
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
ANTIBODIES
BIOCHEMICAL REACTION KINETICS
SPECIFICITY
KERATIN
LABELLING
ANIMAL CELLS
AUTORADIOGRAPHY
CHROMATOGRAPHY
DERMATITIS
EPIDERMIS
EXTRACELLULAR SPACE
FLUORESCENCE
IODINE 125
MICE
MOLECULAR WEIGHT
PATIENTS
TWO-DIMENSIONAL CALCULATIONS
ANIMAL TISSUES
ANIMALS
BETA DECAY RADIOISOTOPES
BODY
DAYS LIVING RADIOISOTOPES
DISEASES
ELECTRON CAPTURE RADIOISOTOPES
EPITHELIUM
INTERMEDIATE MASS NUCLEI
IODINE ISOTOPES
ISOTOPES
KINETICS
LUMINESCENCE
MAMMALS
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
ORGANS
PROTEINS
RADIOISOTOPES
REACTION KINETICS
RODENTS
SCLEROPROTEINS
SEPARATION PROCESSES
SKIN
SKIN DISEASES
SPACE
TISSUES
VERTEBRATES
550201* - Biochemistry- Tracer Techniques