Purification of a Ran-interacting protein that is required for protein import into the nucleus.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 91 ·No. 21 ·1994-10-11 ·Pages 10212-6

Moore MS, Blobel G

Abstract

Previously we reported the isolation of two cytosolic fractions (A and B) from Xenopus ovary that are required sequentially to support protein import into the nuclei of digitonin-permeabilized cells. Fraction A is required for recognition of the nuclear localization sequence and targeting to the nuclear envelope, whereas fraction B is required for the subsequent translocation of the bound substrate into the nucleus. The first protein required for fraction B activity to be purified was the small GTPase Ran (ras-related nuclear protein). Here we report the purification of the second (and final) protein required for fraction B activity. By SDS/PAGE, the purified protein appeared as a single band with an apparent molecular mass of 10 kDa, but the native protein fractionated upon gel filtration chromatography with an apparent size of 30 kDa. Peptide sequence analysis indicated that the purified protein was highly homologous to a previously identified human protein of unknown function called placental protein 15 (pp15) and to the predicted protein product of a yeast open reading frame from Saccharomyces cerevisiae.

MeSH Terms

Amino Acid Sequence Animals Carrier Proteins/isolation & purification,metabolism Cell Nucleus/metabolism Chromatography, DEAE-Cellulose Chromatography, Gel Chromatography, Ion Exchange Cytosol/metabolism Female GTP-Binding Proteins/metabolism Kinetics Liver/metabolism Molecular Sequence Data Nuclear Proteins/isolation & purification,metabolism Nucleocytoplasmic Transport Proteins Ovary/metabolism Rats Rats, Inbred BUF Saccharomyces cerevisiae Proteins Xenopus Proteins Xenopus laevis ran GTP-Binding Protein

Chemicals

Carrier Proteins NTF2 protein, S cerevisiae Nuclear Proteins Nucleocytoplasmic Transport Proteins Ran-interacting protein p10, Xenopus Saccharomyces cerevisiae Proteins Xenopus Proteins GTP-Binding Proteins ran GTP-Binding Protein

Authors & Affiliations

2 authors, click to expand affiliations / ORCID

Moore M S

Laboratory of Cell Biology, Howard Hughes Medical Institute, Rockefeller University, New York, NY 10021.

Blobel G

References (21)

21 references, click to expand

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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.91.21.10212

Published

1994-10-11

Pages

10212-6

Language

English

Region

United States

NLM ID

7505876

PMCID

PMC44988

Subset

External Links

PubMed source DOI: 10.1073/pnas.91.21.10212 PMC Full Text View journal details

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Purification of a Ran-interacting protein that is required for protein import into the nucleus.

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