Hhat is a palmitoylacyltransferase with specificity for N-palmitoylation of Sonic Hedgehog Journal Article
| Authors: | Buglino, J. A.; Resh, M. D. |
| Article Title: | Hhat is a palmitoylacyltransferase with specificity for N-palmitoylation of Sonic Hedgehog |
| Abstract: | Palmitoylation of Sonic Hedgehog (Shh) is critical for effective long- and short-range signaling. Genetic screens uncovered a potential palmitoylacyltransferase (PAT) for Shh, Hhat, but the molecular mechanism of Shh palmitoylation remains unclear. Here, we have developed and exploited an in vitro Shh palmitoylation assay to purify Hhat to homogeneity. We provide direct biochemical evidence that Hhat is a PAT with specificity for attaching palmitate via amide linkage to the N-terminal cysteine of Shh. Other palmitoylated proteins (e.g. PSD95 and Wnt) are not substrates for Hhat, and Porcupine, a putative Wnt PAT, does not palmitoylate Shh. Neither autocleavage nor cholesterol modification is required for Shh palmitoylation. Both the Shh precursor and mature protein are N-palmitoylated by Hhat, and the reaction occurs during passage through the secretory pathway. This study establishes Hhat as a bona fide Shh PAT and serves as a model for understanding how secreted morphogens are modified by distinct PATs. © 2008 by The American Society for Biochemistry and Molecular Biology, Inc. |
| Keywords: | signal transduction; protein expression; unclassified drug; genetics; nonhuman; polymerase chain reaction; animal cell; animal; metabolism; animals; confocal microscopy; green fluorescent protein; cell line; sonic hedgehog protein; hedgehog proteins; morphogenesis; enzyme activity; transfection; cercopithecus aethiops; cos cells; hybrid protein; genetic transfection; iodine 125; isotope labeling; substrate specificity; recombinant protein; cell strain cos1; cercopithecus; cholesterol; enzyme specificity; erinaceidae; fatty acid; enzyme purification; synthesis; acyltransferase; complementary dna; in vitro; cell lysate; immunofluorescence microscopy; polyacrylamide gel electrophoresis; amides; amide; phospholipids; tritium; phospholipid; palmitoylation; lipoylation; radioiodination; acyltransferases; amide linkages; autocleavage; bio-chemicals; bona fide; genetic screens; mature proteins; molecular mechanisms; morphogens; secretory pathways; terminal cysteines; acyl coenzyme a; aminoacyltransferase; palmitic acid; palmitoylacyltransferase; sodium iodide; palmitic acid derivative; protein palmitoyl acyltransferase, human; protein-palmitoyl acyltransferase, human; shh protein, human; fatty acids, nonesterified; palmitates |
| Journal Title: | Journal of Biological Chemistry |
| Volume: | 283 |
| Issue: | 32 |
| ISSN: | 0021-9258 |
| Publisher: | American Society for Biochemistry and Molecular Biology |
| Date Published: | 2008-08-08 |
| Start Page: | 22076 |
| End Page: | 22088 |
| Language: | English |
| DOI: | 10.1074/jbc.M803901200 |
| PUBMED: | 18534984 |
| PROVIDER: | scopus |
| PMCID: | PMC2494920 |
| DOI/URL: | |
| Notes: | --- - "Cited By (since 1996): 38" - "Export Date: 17 November 2011" - "CODEN: JBCHA" - "Source: Scopus" |
