Advertisement
JBC

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Originally published In Press as doi:10.1074/jbc.M212418200 on January 7, 2003

J. Biol. Chem., Vol. 278, Issue 12, 10649-10656, March 21, 2003
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
278/12/10649    most recent
M212418200v1
Right arrow Submit a Letter to Editor
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowRequest Permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Kozono, D.
Right arrow Articles by Kitagawa, Y.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Kozono, D.
Right arrow Articles by Kitagawa, Y.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

Functional Expression and Characterization of an Archaeal Aquaporin
AqpM FROM METHANOTHERMOBACTER MARBURGENSIS*

David KozonoDagger §, Xiaodong Ding§, Ikuko Iwasaki, Xianying Meng||, Yoichi Kamagata||, Peter AgreDagger **, and Yoshichika KitagawaDagger Dagger

From the Dagger  Departments of Biological Chemistry and Medicine, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205-2185,  Biotechnology Institute, Akita Prefectural University, Ogata, 010-0444, and || Microbial and Genetic Resources Research Group, Research Institute of Biological Resources, National Institute of Advanced Industrial Science and Technology, Central 6, Higashi 1-1-1, Tsukuba, Ibaraki 305-8566, Japan

Researchers have described aquaporin water channels from diverse eubacterial and eukaryotic species but not from the third division of life, Archaea. Methanothermobacter marburgensis is a methanogenic archaeon that thrives under anaerobic conditions at 65 °C. After transfer to hypertonic media, M. marburgensis sustained cytoplasmic shrinkage that could be prevented with HgCl2. We amplified aqpM by PCR from M. marburgensis DNA. Like known aquaporins, the open reading frame of aqpM encodes two tandem repeats each containing three membrane-spanning domains and a pore-forming loop with the signature motif Asn-Pro-Ala (NPA). Unlike other known homologs, the putative Hg2+-sensitive cysteine was found proximal to the first NPA motif in AqpM, rather than the second. Moreover, amino acids distinguishing water-selective homologs from glycerol-transporting homologs were not conserved in AqpM. A fusion protein, 10-His-AqpM, was expressed and purified from Escherichia coli. AqpM reconstituted into proteoliposomes was shown by stopped-flow light scattering assays to have elevated osmotic water permeability (Pf = 57 µm·s-1 versus 12 µm·s-1 of control liposomes) that was reversibly inhibited with HgCl2. Transient, initial glycerol permeability was also detected. AqpM remained functional after incubations at temperatures above 80 °C and formed SDS-stable tetramers. Our studies of archaeal AqpM demonstrate the ubiquity of aquaporins in nature and provide new insight into protein structure and transport selectivity.

* This work was supported by a grant-in-aid (to Y. K.) from the Japanese Society for Promotion of Science Postdoctoral Fellowship P00208 for foreign researcher and grants from the National Institutes of Health and the Human Frontier Science Program.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

§ Both authors contributed equally to this work.

** To whom correspondence may be addressed: Dept. Biological Chemistry, The Johns Hopkins School of Medicine, 725 N. Wolfe St., Baltimore, MD 21212-2185. Tel.: 410-955-7049; Fax: 410-955-3149; E-mail: pagre@jhmi.edu.

Dagger Dagger To whom correspondence may be addressed: Biotechnology Institute, Akita Prefectural University, Ogata, 010-0044, Japan. Tel.: 81-185-45-3930; Fax: 81-185-45-2678; E-mail: kitagawa@agri.akita-pu.ac.jp.

Copyright © 2003 by The American Society for Biochemistry and Molecular Biology, Inc.
Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati    What's this?


This article has been cited by other articles:


Home page
 J. Biol. Chem.Home page
G. P. Bienert, A. L. B. Moller, K. A. Kristiansen, A. Schulz, I. M. Moller, J. K. Schjoerring, and T. P. Jahn
Specific Aquaporins Facilitate the Diffusion of Hydrogen Peroxide across Membranes
J. Biol. Chem., January 12, 2007; 282(2): 1183 - 1192.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
S. Gazzarrini, M. Kang, S. Epimashko, J. L. Van Etten, J. Dainty, G. Thiel, and A. Moroni
Chlorella virus MT325 encodes water and potassium channels that interact synergistically
PNAS, April 4, 2006; 103(14): 5355 - 5360.
[Abstract] [Full Text] [PDF]

Home page
 Appl. Environ. Microbiol.Home page
M. J. Daniels, M. R. Wood, and M. Yeager
In Vivo Functional Assay of a Recombinant Aquaporin in Pichia pastoris
Appl. Envir. Microbiol., February 1, 2006; 72(2): 1507 - 1514.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
J. K. Lee, D. Kozono, J. Remis, Y. Kitagawa, P. Agre, and R. M. Stroud
Structural basis for conductance by the archaeal aquaporin AqpM at 1.68 A
PNAS, December 27, 2005; 102(52): 18932 - 18937.
[Abstract] [Full Text] [PDF]

Home page
 Plant Cell PhysiolHome page
M. I. Anderca, S. Suga, T. Furuichi, K. Shimogawara, M. Maeshima, and S. Muto
Functional Identification of the Glycerol Transport Activity of Chlamydomonas reinhardtii CrMIP1
Plant Cell Physiol., September 15, 2004; 45(9): 1313 - 1319.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
 All ASBMB Journals   Molecular and Cellular Proteomics 
 Journal of Lipid Research   ASBMB Today 
Copyright © 2003 by the American Society for Biochemistry and Molecular Biology.
Advertisement
spacer
Advertisement
Advertisement