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Mutations in LRRK2
encoding leucine-rich repeat kinase 2 are thus far the most frequent genetic
cause associated with autosomal dominant and idiopathic Parkinson™s
disease (PD). To examine whether LRRK2 is directly associated with
neuropathology of PD and other related disorders, Xiongwei Zhu and his
collaborators analyzed LRRK2 in brains of patients affected by PD and
dementia with Lewy bodies (DLB) using highly specific antibodies to
LRRK2. In the study described herein (Zhu X. et al. 2006 Molecular
Neurodegeneration 1: 17), the authors showed that anti-LRRK2 antibodies
strongly labeled brainstem and cortical Lewy bodies, the pathological
hallmarks of PD and DLB, respectively. Interestingly, the
immunocytochemical profile of LRRK2 varied with different antibodies
depending upon specific antigenic sites along the LRRK2 protein.
Although all tested antibodies were found immunoreactive in western
blot, only antibodies raised against the N-terminal and C-terminal
regions of LRRK2 were reactive with tissue sections.
Abgent has a collection of 15 antibodies against different regions of
LRRK2. In this study, the PARK8 (LRRK2) Center antibody (product Cat.
No. AP7099b) is raised using a peptide corresponding to the central
region of LRRK2 (amino acid 1246-1265, see Ab2 in Figure 1). Three other
antibodies (Ab1, Ab3 and Ab4) obtained from a commercial source were
also used in this study to monitor epitope exposure within different
regions of LRRK2. Recombinant LRRK2 expressed in human embryonic kidney
HEK293T and human neuroblastoma M17 cells was specifically recognized in
western blot by all four LRRK2 antibodies (Figure 2). However, Ab2 did
not label Lewis bodies by immunohistochemistry staining of LRRK2 in
brainstem sections (data not shown).
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Figure 1: Four antibodies raised against sequences
corresponding to various regions shown on the schematic diagram of LRRK2
protein were used in western blot and immunohistochemistry analysis. The
Ab2 antibody corresponds to Abgent product Cat. No. AP7099b LRRK2
antibody, which binds within the leucin-rich repeats. LRR, leucine-rich
repeats; ROC, Ras of complex protein; COR, C-terminal of ROC; MAPKKK,
mitogen-activated protein kinase kinase kinase; WD40, WD40 repeats.
Figure 2: Western blotting analysis of cell lysates from HEK293T and M17
cells transfected with a LRRK2-expressing plasmid (+). LRRK2 is not
detected in non-transfected cells (-).
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