Tissue vitronectin in normal adult human dermis is non-covalently bound to elastic tissue.
Vitronectin is a multifunctional human plasma glycoprotein that is also found in constant association with elastic tissue fibers in normal adults. We have investigated the nature of the association of vitronectin with elastic tissue, and compared it to that of other elastic fiber-associated proteins, namely fibrillin and amyloid P component. Samples of normal human dermis were incubated with a variety of extraction agents, including high molar salt solution, non-ionic detergent (Nonidet P-40), the reducing agents dithiothreitol or 2-mercaptoethanol, and the chaotropic agents sodium dodecyl sulfate or guanidine hydrochloride. Vitronectin purified from serum typically migrates as two bands of 75 and 65 kD. By contrast, immunoblotting studies of residual dermal material after extraction with the various agents revealed only lower molecular weight (58, 50, 42, 35, and 27 kD) anti-vitronectin reactive bands. Although these bands may represent degradation products of vitronectin generated as a result of the extraction procedure, we cannot exclude the possibility that tissue vitronectin is distinct from plasma vitronectin. Anti-vitronectin reactive polypeptides co-migrating with the 58-, 50-, and 42-kD bands were solubilized following extraction with sodium dodecyl sulfate or guanidine hydrochloride, but not with the other extraction agents. Immunofluorescence studies using residual dermal material after extraction with guanidine hydrochloride demonstrated a marked reduction in elastic fiber staining intensity with anti-vitronectin and anti-amyloid P component, but not with anti-fibrillin. Thus the majority, if not all of dermal vitronectin, is, like amyloid P component, non-covalently associated with, and not an integral constituent of, elastic fibers.
Hintner
H
H
Department of Dermatology, University of Innsbruck, Austria.
Dahlbäck
K
K
Dahlbäck
B
B
Pepys
M B
MB
Breathnach
S M
SM
D001798:Blood Proteins / Q000032:analysis*; D004547:Elastic Tissue / Q000737:chemistry*; D005455:Fluorescent Antibody Technique; D006023:Glycoproteins / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification; D006801:Humans; D008970:Molecular Weight; D000683:Serum Amyloid P-Component / Q000032:analysis / Q000302:isolation & purification; D012867:Skin / Q000737:chemistry*; D019096:Vitronectin
D001798:Blood Proteins; D006023:Glycoproteins; D000683:Serum Amyloid P-Component; D019096:Vitronectin