HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 276, Issue 29, 27111-27119, July 20, 2001

This Article Full Text Full Text (PDF) All Versions of this Article: 276/29/27111    most recent M101410200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Omori, K. Articles by Akatsuka, H. Search for Related Content PubMed PubMed Citation Articles by Omori, K. Articles by Akatsuka, H. Social Bookmarking                      What's this?

Serratia ATP-binding Cassette Protein Exporter, Lip, Recognizes a Protein Region Upstream of the C Terminus for Specific Secretion^*

Kenji Omori^§, Akiko Idei^¶, and Hiroyuki Akatsuka^¶

From the  Discovery Research Laboratory, Tanabe Seiyaku Co., Ltd., 2-50, Kawagishi-2-chome, Toda, Saitama 335-8505, and the ^¶ Discovery Research Laboratory, Tanabe Seiyaku Co., Ltd., 16-89, Kashima-3-chome, Yodogawa-ku, Osaka 532-8505, Japan

Serratia marcescens ATP-binding cassette (ABC) exporter, the Lip system, secretes lipase (LipA_SM), metalloproteases, and a cell surface layer protein homologue but not a heme acquisition protein, HasA (HasA_SM). Secretion of HasA_SM is limited to the Has_SM system. However, HasA proteins from Pseudomonas fluorescens (HasA_PF) and Pseudomonas aeruginosa were exported through the Lip and Has_SM systems. To investigate the specificity in Lip exporter-mediated secretion, secretion analysis was performed using chimeras containing the HasA_PF and HasA_SM sequences. The segment Val-Ala-Leu (designated R1 to R3 sites), which is present close to the C terminus of HasA_PF but not HasA_SM, was revealed to be involved in the substrate specificity of the Lip exporter. Introduction of amino acid substitutions into the R1-R5 region demonstrated that R1, R3, R4, and R5 sites require some specific amino acid residues for Lip-mediated secretion. The amino acid sequence of the region was conserved considerably among the proteins secreted by the Lip exporter. On the contrary, the region was not related to HasA secretion through the Has_SM system. Interestingly, a typical C-terminal motif, so far regarded as a secretion signal, was not necessary for secretion through either the Lip or the Has_SM exporter. In LipA_SM secretion via the Lip system, the typical C-terminal motif was not essential either, but the presence of a sequence similar to Val-Ala-Leu and its location from the C terminus greatly affect the secretion level. Secretion analyses using hybrid exporters and competitors exhibited that the R1-R5 region was recognized by an ABC protein of the Lip exporter, LipB, and that the mutations aborting Lip-mediated secretion in the region resulted in a loss of the affinity to LipB. Thus, a determinant within the secretory protein for Lip-mediated secretion was fully defined.

CiteULike

Complore

Connotea

Del.icio.us

Digg

Reddit

Technorati

This article has been cited by other articles:

				K. Kuwahara, C. Angkawidjaja, H. Matsumura, Y. Koga, K. Takano, and S. Kanaya Importance of the Ca2+-binding sites in the N-catalytic domain of a family I.3 lipase for activity and stability Protein Eng. Des. Sel., December 1, 2008; 21(12): 737 - 744. [Abstract] [Full Text] [PDF]