Cleavage without anchor addition accompanies the processing of a nascent protein to its glycosylphosphatidylinositol-anchored form.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 92 ·No. 5 ·1995-02-28 ·Pages 1550-4

Maxwell SE, Ramalingam S, Gerber LD, Udenfriend S

Abstract

Rough microsomal membranes from most mammalian cells, in the presence of a translation system, process nascent proteins with appropriate COOH-terminal signal peptides to their mature glycosylphosphatidylinositol (GPI)-linked forms. The present study, using preprominiplacental alkaline phosphatase as substrate, shows that as much as 10% of the mature product is cleaved correctly but is not linked to GPI. Some of the factors that influence the relative proportions of GPI linked to free mini-placental alkaline phosphatase are the amounts of GPI in the cells and the amino acid substituent at the omega site of the nascent protein. A mechanism for explaining cleavage both with and without GPI addition is presented, which supports a transamidase type of enzyme as the catalyst.

MeSH Terms

Alkaline Phosphatase/metabolism Animals CHO Cells Cricetinae Glycosylphosphatidylinositols/metabolism HeLa Cells Humans In Vitro Techniques Microsomes/metabolism Molecular Weight Placenta/enzymology Protein Biosynthesis Protein Processing, Post-Translational Proteins/metabolism

Chemicals

Glycosylphosphatidylinositols Proteins phospholipase A2-activating protein Alkaline Phosphatase

Authors & Affiliations

4 authors, click to expand affiliations / ORCID

Maxwell S E

Roche Institute of Molecular Biology, Roche Research Center, Nutley, NJ 07110-1199.

Ramalingam S

Gerber L D

Udenfriend S

References (18)

18 references, click to expand

Mechanism of action of penicillins: a proposal based on their structural similarity to acyl-D-alanyl-D-alanine.
Proc Natl Acad Sci U S A. 1965 Oct;54(4):1133-41 PMID: 5219821
Structural analysis of glycosylphosphatidylinositol anchors.
Methods Enzymol. 1994;230:418-42 PMID: 8139510
Purification and properties of phosphatidylinositol-specific phospholipase C of Bacillus thuringiensis.
Biochim Biophys Acta. 1980 Jul 14;619(1):48-57 PMID: 7417468
Phase separation of integral membrane proteins in Triton X-114 solution.
J Biol Chem. 1981 Feb 25;256(4):1604-7 PMID: 6257680
gamma-Glutamyl transpeptidase: catalytic, structural and functional aspects.
Mol Cell Biochem. 1981 Sep 25;39:357-68 PMID: 6118826
Processing at the carboxyl terminus of nascent placental alkaline phosphatase in a cell-free system: evidence for specific cleavage of a signal peptide.
Proc Natl Acad Sci U S A. 1989 Jan;86(1):22-6 PMID: 2911571
Selectivity of the cleavage/attachment site of phosphatidylinositol-glycan-anchored membrane proteins determined by site-specific mutagenesis at Asp-484 of placental alkaline phosphatase.
Proc Natl Acad Sci U S A. 1990 Jan;87(1):157-61 PMID: 2153284
Selectivity at the cleavage/attachment site of phosphatidylinositol-glycan anchored membrane proteins is enzymatically determined.
Proc Natl Acad Sci U S A. 1990 Oct;87(20):7939-43 PMID: 1700420
Glycophospholipid membrane anchor attachment. Molecular analysis of the cleavage/attachment site.
J Biol Chem. 1991 Jan 15;266(2):1250-7 PMID: 1824699
Biosynthesis of phosphatidylinositol glycan-anchored membrane proteins. Design of a simple protein substrate to characterize the enzyme that cleaves the COOH-terminal signal peptide.
J Biol Chem. 1991 Mar 5;266(7):4464-70 PMID: 1999429
Functional analysis of T-cell mutants defective in the biosynthesis of glycosylphosphatidylinositol anchor. Relative importance of glycosylphosphatidylinositol anchor versus N-linked glycosylation in T-cell activation.
J Biol Chem. 1991 Dec 5;266(34):23175-84 PMID: 1835975
Biosynthesis of phosphatidylinositol-glycan (PI-G)-anchored membrane proteins in cell-free systems: cleavage of the nascent protein and addition of the PI-G moiety depend on the size of the COOH-terminal signal peptide.
Proc Natl Acad Sci U S A. 1992 Feb 15;89(4):1350-3 PMID: 1531539
Biosynthesis of phosphatidylinositol-glycan (PI-G)-anchored membrane proteins in cell-free systems: PI-G is an obligatory cosubstrate for COOH-terminal processing of nascent proteins.
Proc Natl Acad Sci U S A. 1992 Jun 1;89(11):4982-5 PMID: 1594603
Phosphatidylinositol glycan (PI-G) anchored membrane proteins. Amino acid requirements adjacent to the site of cleavage and PI-G attachment in the COOH-terminal signal peptide.
J Biol Chem. 1992 Jun 15;267(17):12168-73 PMID: 1601882
Evidence that the putative COOH-terminal signal transamidase involved in glycosylphosphatidylinositol protein synthesis is present in the endoplasmic reticulum.
Proc Natl Acad Sci U S A. 1993 May 1;90(9):3973-7 PMID: 8387204
The structure, biosynthesis and function of glycosylated phosphatidylinositols in the parasitic protozoa and higher eukaryotes.
Biochem J. 1993 Sep 1;294 ( Pt 2):305-24 PMID: 8373346
Correction of the class H defect in glycosylphosphatidylinositol anchor biosynthesis in Ltk- cells by a human cDNA clone.
J Biol Chem. 1993 Oct 5;268(28):20733-6 PMID: 8407896
Isolation of rat liver plasma membrane fragments in isotonic sucrose.
Methods Enzymol. 1974;31:90-102 PMID: 4370714

Article Info

Journal

Proceedings of the National Academy of Sciences of the United States of America

Abbr.

Proc Natl Acad Sci U S A

ISSN

0027-8424

DOI

10.1073/pnas.92.5.1550

Published

1995-02-28

Pages

1550-4

Language

English

Region

United States

NLM ID

7505876

PMCID

PMC42557

Subset

External Links

PubMed source DOI: 10.1073/pnas.92.5.1550 PMC Full Text View journal details

Analysis Services

Cleavage without anchor addition accompanies the processing of a nascent protein to its glycosylphosphatidylinositol-anchored form.

Contact

WeChat Official Account