A tetrameric porin limits the cell wall permeability of Mycobacterium smegmatis.
Mycobacteria protect themselves with an outer lipid bilayer, which is the thickest biological membrane hitherto known and has an exceptionally low permeability rendering mycobacteria intrinsically resistant against many antibiotics. Pore proteins mediate the diffusion of hydrophilic nutrients across this membrane. Electron microscopy revealed that the outer membrane of Mycobacterium smegmatis contained about 1000 protein pores per microm(2), which are about 50-fold fewer pores per microm(2) than in Gram-negative bacteria. The projection structure of the major porin MspA of M. smegmatis was determined at 17 A resolution. MspA forms a cone-like tetrameric complex of 10 nm in length with a single central pore. Thus, MspA is drastically different from the trimeric porins of Gram-negative bacteria and represents a new class of channel proteins. The formation of MspA micelles indicated that the ends of MspA have different hydrophobicities. Oriented insertion of MspA into membranes was demonstrated in lipid bilayer experiments, which revealed a strongly asymmetrical voltage gating of MspA channels at -30 mV. The length of MspA is sufficient to span the outer membrane and contributes in combination with the tapering end of the pore and the low number of pores to the low permeability of the cell wall of M. smegmatis for hydrophilic compounds.
Max-Planck-Institut für Biochemie, Abteilung Molekulare Strukturbiologie, Am Klopferspitz 18a, D-82152 Martinsried, Germany.
Harald
H
Engelhardt
Christian
C
Heinz
Michael
M
Niederweis
D002473:Cell Wall / Q000378:metabolism* / Q000648:ultrastructure; D002850:Chromatography, Gel; D007091:Image Processing, Computer-Assisted; D008565:Membrane Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000648:ultrastructure; D008854:Microscopy, Electron; D020102:Mycobacterium smegmatis / Q000502:physiology*; D010539:Permeability; D018272:Porins / Q000737:chemistry* / Q000302:isolation & purification / Q000648:ultrastructure*; D011487:Protein Conformation
D008565:Membrane Proteins; D018272:Porins; C121838:mspA protein, Mycobacterium smegmatis