Amyloid P component binds to keratin bodies in human skin and to isolated keratin filament aggregates in vitro.
Dermal keratin bodies, consisting mainly of keratin intermediate filament aggregates (KIFA) coated with IgM anti-KIF autoantibodies, are present in normal human skin and occur in increased quantities in certain skin diseases. Keratin bodies are normally rapidly removed, but in primary localized cutaneous amyloidosis (PLCA) they are converted by an unknown mechanism to amyloid. Amyloid P component (AP), a glycoprotein identical to, and derived from, the normal plasma protein serum amyloid P component (SAP), is present in all forms of amyloid including PLCA. We investigated the interaction between SAP, keratin bodies, and KIFA. Immunofluorescence staining of normal skin using fluoresceinated anti-SAP and rhodamine-conjugated anti-IgM, or AE-1/AE-3 anti-keratin antibodies followed by Texas Red-conjugated anti-mouse immunoglobulin, showed that 52% +/- 4 (mean +/- sem, n = 6) of keratin bodies bound anti-SAP. Similar findings were present in a biopsy from a patient with lichen planus. Isolated KIFA, prepared by 8M urea extraction of normal human epidermis or cultured keratinocytes, were preincubated with normal human serum as a source of SAP and then stained with fluoresceinated anti-SAP. Bright fluorescence seen when the incubation medium contained Ca++ was absent in the presence of ethylenediamine tetraacetic acid. Specific Ca++-dependent binding of SAP to KIFA was confirmed using immunoblotting. Binding of SAP to KIFA did not prevent their degradation following exposure to trypsin or alpha-chymotrypsin. Similarly, partial enzymatic digestion of KIFA did not abrogate their ability to bind SAP. Our findings, that SAP is associated with keratin bodies in skin and exhibits Ca++-dependent binding to KIFA in vitro, identify keratin filaments as a newly recognized ligand for SAP.
Hintner
H
H
Department of Medicine, (Dermatology), Charing Cross and Westminister Medical School, London, U.K.
Booker
J
J
Ashworth
J
J
Auböck
J
J
Pepys
M B
MB
Breathnach
S M
SM
D052160:Benzothiazoles; D002118:Calcium / Q000502:physiology; D002478:Cells, Cultured; D003224:Congo Red; D003599:Cytoskeleton / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007382:Intermediate Filaments / Q000378:metabolism*; D007633:Keratins / Q000378:metabolism*; D000683:Serum Amyloid P-Component / Q000378:metabolism*; D012867:Skin / Q000033:anatomy & histology / Q000378:metabolism*; D013194:Staining and Labeling; D013844:Thiazoles; D014357:Trypsin / Q000378:metabolism
D052160:Benzothiazoles; D000683:Serum Amyloid P-Component; D013844:Thiazoles; C009462:thioflavin T; D003224:Congo Red; D007633:Keratins; D014357:Trypsin; D002118:Calcium