Scripps Institute CMT research

[Guest guest]

(Scripps Institute also found the first case of Spontaneous X CMT - this

research news article mentions CMT ~ Gretchen)

Study at TSRI links dozens of proteins to several rare muscle and nerve

degeneration diseases

La Jolla, CA. September 2, 2003--A team of scientists at The Scripps

Research Institute (TSRI) has identified more than 50 previously unknown

proteins and associates several of them with rare human muscle and nerve

degeneration diseases. The team is publishing their findings this week

in the journal Science. Led by TSRI Professors Larry Gerace and R.

Yates III, the team used a technique called subtractive proteomics to

identify 62 new proteins in the inner

nuclear membrane of the human cell. The team demonstrated that 23 of

these proteins are linked with strong probability to 14 rare

muscle-wasting diseases such as congenital muscular dystrophy,

Limb-Girdle muscular dystrophy, and spinal muscular atrophy, and several

forms of the neurodegenerative Charcot-Marie-Tooth disease.

Knowing the proteins that may cause or contribute to these diseases is a

first step in the long process of looking for ways to detect, prevent,

or treat them.

This study has the potential to clarify a significant number of the

more than 300 human dystrophies for which a causative gene has

not been identified.

" To understand how these diseases happen, we need to understand more

about the players--the network of interlinked proteins, " says Gerace.

Muscular Dystrophies and the Nuclear Membrane

Many rare but devastating diseases have been linked to the inner

nuclear membrane, which lines the nuclear envelope compartmentalizing

the cell's genetic material or DNA. On the

inner surface of the membrane is a structure referred to as the

lamina. The lamina is important for maintaining the shape and size of

the nucleus. It also contributes to the specialized functions

of different human cells, for example, enabling muscle cells to

perform their particular functions and brain cells to perform theirs.

The lamina is largely composed of proteins called lamins, which

are like bricks that form a scaffold-like structure for the nucleus.

The lamina also contains membrane proteins that dock at the lamins.

" There have been a number of human muscular and neuronal

dystrophies that have been linked to [these] proteins, " says Gerace.

" When certain lamins and inner membrane proteins are mutated, they cause

disease. "

Because of this link between lamina proteins and disease, scientists

would like to know the identity of all the proteins in the lamina, and

previous studies have identified about 20 lamina proteins.

In their current study, Gerace and Yates used a technique called

subtractive proteomics to identify 62 more candidate human nuclear

membrane proteins.

In the study, TSRI Postdoctoral Fellow Schirmer demonstrates that

the genes encoding 23 of these candidate human nuclear membrane proteins

are in regions of the genome that have already been implicated in 14

muscle- and neuro-degenerative diseases.

However, many of these regions have hundreds of genes in them; so the

identification of these disease gene candidates should greatly focus

identification of the culprits.

" It's highly likely that some of these diseases will be due to [the

newly identified] nuclear envelope proteins, " says Gerace. " This is

a pretty big step forward. "

The Power of Subtractive Proteomics

Where " genomics " maps the DNA sequence and seeks to identify all the

genes in an organism, " proteomics " takes a step further by asking where

and when those genes are actually expressed as proteins.

One of the most important techniques emerging for proteomics

studies is humbly referred to as MudPIT--Multidimensional Protein

Identification Technology-which Yates has pioneered in the last

few years. Using this technique, scientists like Yates are able to

analyze and identify an enormous number of proteins in a complex

mixture.

MudPIT basically combines liquid chromatography (which is like a

molecular " sieve " that separates a complex mixture into its

component parts) with tandem mass spectrometry (which identifies the

components based on their masses). The instrument detects these masses

and uses sophisticated software to identify thousands of separate

proteins.

But MudPIT alone was not enough in this case, because the inner

nuclear membrane is in contact with other structures of the cell

and cannot be isolated without contaminating material. Identifying

which proteins are from the inner nuclear membrane and which are

contaminants presented a huge problem.

So the team used a simple subtractive technique to deal with this.

They analyzed the nuclear membrane components with contaminants,

containing 2,071 different proteins, and subtracted out the separately

isolated contaminants, which accounted for more than 40 percent of the

membrane proteins. From this list they were able to apply computational

methods to limit the final list of new human nuclear membrane proteins

to 62.

The scientists then took eight of these proteins at random and

demonstrated that they all indeed targeted to the nuclear membrane.

The new nuclear membrane proteins identified in this study map

to chromosomal regions where the following dystrophies have been

localized:

Congenital Ptosis, hereditary type 1

Charcot-Marie-Tooth Disease 2A

Congenital Muscular Dystrophy 1B

Limb-Girdle Muscular Dystrophy 2B

Charcot-Marie-Tooth Disease 2A

Facioscapulohumeral Muscular Dystrophy (FSH)

Spinal Muscular Atrophy, Types 1, 2, and 3

Limb-Girdle Muscular Dystrophy 1A/1B

Arthogryposis: neurogenic, mild

Blepharophimosis 2

Charcot-Marie-Tooth Disease 2A

Distal Arthrogryposis, type 2B

Congenital Fibrosis of Extraocular Muscles 1

Distal Myopathy

The article, " Nuclear Membrane Proteins With Potential Disease

Links Found By Subtractive Proteomics " was authored by C.

Schirmer, ce Florens, Tinglu Guan, R. Yates III, and

Larry Gerace and will appear in the September 5, 2003 issue of

the journal Science. See: http://www.sciencemag.org.