Crystal structure of cathepsin X: a flip-flop of the ring of His23 allows carboxy-monopeptidase and carboxy-dipeptidase activity of the protease.
BACKGROUND
Cathepsin X is a widespread, abundantly expressed papain-like mammalian lysosomal cysteine protease. It exhibits carboxy-monopeptidase as well as carboxy-dipeptidase activity and shares a similar activity profile with cathepsin B. The latter has been implicated in normal physiological events as well as in various pathological states such as rheumatoid arthritis, Alzheimer's disease and cancer progression. Thus the question is raised as to which of the two enzyme activities has actually been monitored.


RESULTS
The crystal structure of human cathepsin X has been determined at 2.67 A resolution. The structure shares the common features of a papain-like enzyme fold, but with a unique active site. The most pronounced feature of the cathepsin X structure is the mini-loop that includes a short three-residue insertion protruding into the active site of the protease. The residue Tyr27 on one side of the loop forms the surface of the S1 substrate-binding site, and His23 on the other side modulates both carboxy-monopeptidase as well as carboxy-dipeptidase activity of the enzyme by binding the C-terminal carboxyl group of a substrate in two different sidechain conformations.


CONCLUSIONS
The structure of cathepsin X exhibits a binding surface that will assist in the design of specific inhibitors of cathepsin X as well as of cathepsin B and thereby help to clarify the physiological roles of both proteases.
Guncar
G
G
Department of Biochemistry and Molecular Biology, Jozef Stefan Institute, Ljubljana, 1000, Slovenia.
Klemencic
I
I
Turk
B
B
Turk
V
V
Karaoglanovic-Carmona
A
A
Juliano
L
L
Turk
D
D
D000595:Amino Acid Sequence; D001665:Binding Sites; D002268:Carboxypeptidases / Q000378:metabolism*; D056657:Cathepsin K; D002403:Cathepsins / Q000737:chemistry* / Q000378:metabolism; D018360:Crystallography, X-Ray; D006639:Histidine / Q000737:chemistry*; D006801:Humans; D007700:Kinetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017386:Sequence Homology, Amino Acid
D006639:Histidine; D002268:Carboxypeptidases; D002403:Cathepsins; C534130:CTSK protein, human; D056657:Cathepsin K