title: Penetration of three transmembrane segments of Slc11a1 in lipid bilayers.
abstract: Slc11a1 is a divalent metal cation transporter with 12 putative transmembrane domains (TM) and plays a role in host defense. In present work, we investigated the secondary structure and topology of the peptides associated to Slc11a1-TM2, TM3 and TM4 (wildtype peptides and function-relating mutants) in the phospholipid vesicles (DMPC, DMPG and their mixtures) using circular dichroism, fluorescence spectroscopy and differential scanning calorimetry. We found that TM3 is obviously different in secondary structure and topology from TM2 to TM4 in the lipid membranes. The peptide TM3 is less structured and embedded in the lipid membranes less deeply than TM2 and TM4 at pH 5.5 and 7. The insertion position of TM3 in the lipid membranes is adjusted by pH, more deeply at more acidic pH environment, whereas the locations of TM2 and TM4 in the lipid membranes are less changed with pH. The E139A substitution of TM3 significantly impairs the pH dependence of the buried depth of TM3 and causes a pronounced increase in helicity in all DMPG-containing lipid vesicles at pH 5.5 and 7 and in DMPC at pH 4. In contrast, TM2 and TM4 are similar in topology. The G169D mutation has little effect on the topological arrangement of TM4 in membranes. The property of headgroups of the phospholipids has an effect on the secondary structure and topology of the peptides. All peptides could be structured with more helicity and embedded more deeply in DMPG-containing lipid vesicles than in DMPC membrane at pH 5.5 and 7.
authors.affiliation: College of Chemistry and Chemical Engineering, Qiqihar University, No. 42, Wenhua Street, Qiqihar, PR China. Electronic address: qhy120@sina.com.
authors.forename: Haiyan
authors.initials: H
authors.lastname: Qi
authors.affiliation: College of Chemistry and Chemical Engineering, Qiqihar University, No. 42, Wenhua Street, Qiqihar, PR China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Wang
authors.affiliation: College of Chemistry and Chemical Engineering, Qiqihar University, No. 42, Wenhua Street, Qiqihar, PR China.
authors.forename: Hongtao
authors.initials: H
authors.lastname: Chu
authors.affiliation: College of Chemistry and Chemical Engineering, Qiqihar University, No. 42, Wenhua Street, Qiqihar, PR China.
authors.forename: Wenhua
authors.initials: W
authors.lastname: Wang
authors.affiliation: College of Chemistry and Chemical Engineering, Qiqihar University, No. 42, Wenhua Street, Qiqihar, PR China.
authors.forename: Qidong
authors.initials: Q
authors.lastname: Mao
mesh_terms: D020106:Acrylamide / Q000737:chemistry; D000595:Amino Acid Sequence; D002152:Calorimetry, Differential Scanning; D027682:Cation Transport Proteins / Q000737:chemistry*; D002462:Cell Membrane / Q000737:chemistry*; D002942:Circular Dichroism; D004134:Dimyristoylphosphatidylcholine / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D008051:Lipid Bilayers / Q000737:chemistry*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D050505:Mutant Proteins / Q000737:chemistry; D010455:Peptides / Q000737:chemistry*; D010539:Permeability; D010715:Phosphatidylglycerols / Q000737:chemistry; D010743:Phospholipids / Q000737:chemistry; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D013050:Spectrometry, Fluorescence; D053835:Unilamellar Liposomes / Q000737:chemistry; D000099173:Natural Resistance-associated Macrophage Protein 1
chemical_list: D020106:Acrylamide; D027682:Cation Transport Proteins; D004134:Dimyristoylphosphatidylcholine; D008051:Lipid Bilayers; D050505:Mutant Proteins; D010455:Peptides; D010715:Phosphatidylglycerols; D010743:Phospholipids; D053835:Unilamellar Liposomes; D000099173:Natural Resistance-associated Macrophage Protein 1; C002773:dimyristoylphosphatidylglycerol
keywords: CD; DSC; Fluorescence; Phospholipids; Slc11a1; Transmembrane domains