title: Hydrolytic enzymes of the interfollicular epidermis differ in expression and correlate with the phenotypic difference observed between light and dark skin.
abstract: Degradation of melanosomes in light skin (LS, i.e. phototype I/II) appears to occur more rapidly than dark skin (DS, i.e. phototype IV/V). Hydrolytic enzymes known to reside and be expressed in a differential pattern within the interfollicular epidermis are implicated in playing a role in epidermal differentiation and potentially melanosome degradation. The aim of this present study was to evaluate the differential expression of hydrolytic enzymes that may correlate with physiological and phenotypic differences seen between DS and LS. Expression of six hydrolytic enzymes was confirmed by microarray analysis of the suprabasal epidermis from LS and DS. Specific lysosomal hydrolases identified by microarray analysis were analyzed by indirect immunofluorescence (IIF) and immunoblot analysis. Immunogold electron microscopy (IEM) was completed to visualize cellular expression of the hydrolytic enzyme cathepsin L2 (Cath L2) and biochemical assay was performed to ascertain Cath L2 activity. Immunoblotting of light and dark epidermal lysates demonstrated that of the six enzymes initially analyzed, both prostatic acid phosphatase (ACPP) and Cath L2 were reproducibly upregulated in DS and LS, respectively. IIF and IEM analyses of Cath L2 in tissue confirmed this differential expression. Biochemical analysis of Cath L2 in light and dark epidermal lysates displays increased activity of Cath L2 in LS. The results of this study confirm differential expression of ACPP and Cath L2 in DS and LS at gene and protein level. Additionally, Cath L2 displays increased activity in LS-derived epidermal lysates. This study indentified two acid hydrolases that may play a role in melanosome degradation and pigment processing.
authors.affiliation: College of Pharmacy, University of Cincinnati, Cincinnati, Ohio, USA.
authors.forename: Jody P
authors.initials: JP
authors.lastname: Ebanks
authors.forename: Amy
authors.initials: A
authors.lastname: Koshoffer
authors.forename: R Randall
authors.initials: RR
authors.lastname: Wickett
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Hakozaki
authors.forename: Raymond E
authors.initials: RE
authors.lastname: Boissy
mesh_terms: D000135:Acid Phosphatase; D002403:Cathepsins / Q000378:metabolism; D002478:Cells, Cultured; D003546:Cysteine Endopeptidases / Q000378:metabolism; D004817:Epidermis / Q000201:enzymology*; D052816:Foreskin; D006801:Humans; D006868:Hydrolysis; D015151:Immunoblotting; D007150:Immunohistochemistry; D007231:Infant, Newborn; D008297:Male; D020460:Melanosomes / Q000201:enzymology* / Q000235:genetics; D008854:Microscopy, Electron; D020411:Oligonucleotide Array Sequence Analysis; D010641:Phenotype; D017027:Protein Tyrosine Phosphatases / Q000378:metabolism*; D012880:Skin Pigmentation / Q000235:genetics*
chemical_list: D000135:Acid Phosphatase; C428439:prostatic acid phosphatase; D017027:Protein Tyrosine Phosphatases; D002403:Cathepsins; D003546:Cysteine Endopeptidases; C117627:CTSV protein, human