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. 2000 Jul 10;150(1):77–88. doi: 10.1083/jcb.150.1.77

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© 2000 The Rockefeller University Press

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).

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Processing of Gas1p and CPY in per mutants. Logarithmically growing wild-type and mutant cells were metabolically labeled with [³⁵S]methionine/cysteine for 5 min at 30°C, followed by a chase for 30 min. From detergent lysates, Gas1p and CPY were immunoprecipitated using monospecific polyclonal antisera. As a control, these proteins were also immunoprecipitated from wild-type lysates pulse-labeled with no chase to indicate the ER forms (lane 1). Proteins are separated on a 10–15% polyacrylamide gradient gel and visualized by fluorography. The positions of preGas1p, ER Gas1p, and Golgi/plasma membrane Gas1p (Golgi/PM) are indicated (A). For CPY, the ER (P1), Golgi apparatus (P2), and mature vacuolar form (CPY) are indicated to the left and the vacuolar underglycosylated forms (-1, -2, -3, -4) to the right of B.