STRUCTURAL BIOLOGY OF MEMBRANE PROTEINS

Release Date:  October 16, 1998

PA NUMBER:  PA-99-004

P.T.

National Institute of General Medical Sciences
National Institute of Arthritis and Musculoskeletal and Skin Diseases 
National Institute of Diabetes and Digestive and Kidney Diseases
National Institute of Environmental Health Sciences
National Institute of Neurological Disorders and Stroke

PURPOSE

The purpose of this program announcement (PA) is to encourage basic
research on the structures of membrane proteins at (or near) atomic
resolution.  Considerable research is on-going in the area of
membrane protein structure and function, particularly with respect
to sequences, topology, and the effects of mutations; however, much
of this work is somewhat speculative in that the interpretations
depend upon the very limited number of structures that have
actually been solved by direct biophysical measurements.  Despite
several recent landmark solutions of membrane protein structures,
there remains a significant gap between the understanding of
membrane proteins and that of their soluble counterparts.  This gap
will likely increase as the facility with which soluble protein
structures can be solved continues to increase.  Therefore, it is
clear that a special effort is needed to promote studies of
membrane protein structures.  An increase in the number of known
membrane protein structures will contribute to an enhanced
understanding of many basic phenomena underlying cellular functions
essential to human health.

HEALTHY PEOPLE 2000

The Public Health Service (PHS) is committed to achieving the
health promotion and disease prevention objectives of "Healthy
People 2000," a PHS led national activity for setting priority
areas. This Program Announcement (PA), Title of PA, is related to
one or more of the priority areas. Potential applicants may obtain
a copy of "Healthy People 2000" at
http://www.crisny.org/health/us/health7.html.

ELIGIBILITY REQUIREMENTS

Applications may be submitted by foreign and domestic, for-profit
and non-profit organizations, public and private, such as
universities, colleges, hospitals, laboratories, units of State and
local governments, and eligible agencies of the Federal government. 
Racial/ethnic minority individuals, women, and persons with
disabilities are encouraged to apply as principal investigators. 
Foreign institutions are not eligible for the SBIR (R43, R44), STTR
(R41, R42), or program project (P01) grant mechanisms.

MECHANISM OF SUPPORT

Support of this program will be through the individual research
project grant (R01), program project grant (P01).  Applicants may
also apply for the Small Business Innovation Research (R43, R44)
and Small Business Technology Transfer (R41, R42) award mechanisms. 
Details of STTR and SBIR mechanisms (i.e., application procedures,
review procedures) are provided in the Omnibus Solicitations of the
NIH, CDC, and FDA for Small Business Innovation Research Grant
Applications (SBIR 98-2) and the Omnibus Solicitation of the
National Institutes of Health for Small Business Technology
Transfer Grant Applications (PHS 98-3).  These documents are
available on the NIH Website: 
http://www.nih.gov/grants/funding/sbir.htm.  Copies may also be
obtained by calling the SBIR/STTR Solicitation Office:
301-206-9385.

Investigators holding active R01, P01, or MERIT (R37) grants to
study membrane associated processes, but who are not currently
supported for work on high resolution structural studies of the
relevant membrane associated proteins, may wish to consider
applying for competing supplemental awards, if they would have at
least one year remaining in the project period at the time of
supplement funding.

All potential applicants are strongly urged to contact the program
staff listed under INQUIRIES for guidance in the areas appropriate
for this program, especially if submission of a program project or
supplemental grant application is being considered

RESEARCH OBJECTIVES

Membrane proteins play a crucial role in many cellular and
physiological processes.  They are essential mediators of material
and information transfer between cells and their environment,
between compartments within cells, and between compartments
comprising the organ systems.  Functionally normal membrane
proteins are vital to health and specific defects are associated
with many known disease states.  Membrane proteins are the targets
of a large number of pharmacologically and toxicologically active
substances and are responsible, in part, for their uptake,
metabolism, and clearance.

Despite the importance of membrane associated proteins, the
knowledge of their high resolution structures and mechanisms of
action has lagged far behind the knowledge of these properties of
proteins in general.  This has resulted from the difficulty of
obtaining x-ray diffraction quality crystals for the membrane-
embedded domains of these proteins and the difficulty of applying
well developed solution NMR methods to the study of most membrane
proteins.  These difficulties have led to a reluctance of many
investigators to pursue high resolution structural studies of
membrane proteins.  However, in the recent past, advances in
methods for crystallization and analysis of proteins by x-ray and
electron diffraction methods, and improvements in NMR methods,
particularly solid-state NMR, have led to new opportunities. 
Further, the solution of crystal structures, once suitable crystals
are obtained has, in many cases, become sufficiently routine, that
crystallization itself is often a major undertaking worthy of
financial support.  The objective of this program announcement is
two-fold:

1) To encourage investigators with interests in membrane associated
systems to pursue high resolution structural studies making use of
these recently developed technologies; and

2) To encourage additional research to further develop methods for
studying the structure of membrane proteins at atomic resolution.
Areas identified as needing specific attention include:

o  Improved methods for over-expression of native and modified
membrane proteins,

o  Improved methods for isolation, purification, and stabilization
of membrane proteins, including the development of new detergents
and non-detergent solubilization agents,

o  Basic research on the physical chemistry of membrane protein
crystallization and the development of new methods for
crystallization and crystal manipulation that could facilitate data
collection,

o  Further development of methods for electron diffraction,
particularly for the production of suitable 2D-crystals,

o  Further development of NMR methods for examining membrane
proteins in their native lipid environments.

The techniques of x-ray or electron diffraction and of NMR
spectroscopy have been emphasized in this announcement, since it is
felt that they show the most promise for producing complete high
resolution information for the largest number of proteins. 
However, funds are also available for research using other methods
that can provide atomic resolution information in selected cases. 
Methods that can elucidate the organization of lipid and detergent
molecules within protein crystalline arrays (e.g., neutron
diffraction) are also of interest.

It is expected that many of the projects will be collaborative
efforts between biochemists and molecular biologists with expertise
in the isolation and characterization of membrane-bound proteins
and biophysicists with expertise in x-ray crystallography, NMR, and
other structural methods.  A major aim of this program announcement
is to stimulate such collaborations.

Membrane protein systems of particular interest to the National
Institute of General Medical Sciences (NIGMS) include:  energy
transducing membranes of mitochondria, chloroplasts, and bacterial
cell membranes involved in electron transport and ATP synthesis;
transporters of ions, substrates, and macromolecules between
intracellular compartments and between the cell and its
environment; enzymes in the synthesis and metabolism of lipids,
membrane-associated and secreted proteins, and glycoconjugates;
cytoskeletal proteins, including those required for intracellular
vesicle transport, cell motility, and cell division; regulators of
cell-cell communication, differentiation, and growth; receptors
relevant to cell cycle regulation, mechanisms of anesthetic action,
and trauma and burn physiology; transporters and enzymes
responsible for the uptake, metabolism, and clearance of drugs, or
in other ways affecting the bioavailability, pharmacokinetics, or
action of drugs; targets of drug action and toxicity, including
targets of naturally occurring toxins and venoms; enzymes involved
in the biosynthesis of natural products.

Membrane protein systems of interest to the National Institute of
Arthritis and Musculoskeletal and Skin Diseases (NIAMS) have
specific relevance to one of the following programmatic areas:
muscle function and disease; bone and cartilage function and
disease; skin function and disease.  Examples of NIAMS interests
are: structural aspects of membrane proteins in muscle disease,
excitation, relaxation, force transduction, cellular homeostasis
and metabolism, regulators of cell-cell communication and
attachment (e.g. costameres, myotendinous and neuromuscular
junctions), ion channels, receptors, transporters and enzymes that
effect muscle function and hypertrophy or atrophy; and structural
aspects of membrane proteins in skin as they are involved in the
establishment of the stratum corneum barrier, epidermal cell-cell
attachment and communication, transmembrane signaling and
transport, and cell movement, including genetic and acquired
diseases of skin in which the membrane protein is defective or
targeted (which may encompass both benign and malignant
hyperproliferative diseases).

Membrane protein systems of particular interest to the National
Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
should have specific relevance to one of the following programmatic
areas: diseases of transport such as cystic fibrosis and
peroxisomal biogenesis disorders; carbohydrate metabolism and its
hormonal control; diabetes mellitus; hormone receptors and signal
transduction; endocrine disorders; normal and abnormal processes of
lipid, protein, amino acid, urea, pyrimidine, metal ion and steroid
metabolism; genetic metabolic disorders.  Proteins should be of
mammalian origin.  Studies on proteins of prokaryotic or lower
eukaryotic origin should be proposed as models for mammalian
systems.  An example of this is the ABC transporter superfamily or
traffic ATPases in bacteria and yeast that serve as models for the
cystic fibrosis transmembrane regulator (CFTR).

Membrane protein systems of particular interest to the National
Institute of Environmental Health Sciences (NIEHS) include those
proteins/enzymes involved in the response of cells to environmental
toxicants.  These proteins/enzymes may include the components of
the stress signaling pathway, ion channels involved in transport of
xenobiotics, i.e., membrane transporters as PgP and MDR, MRP2 ,
transporters and enzymes responsible for the uptake metabolism and
clearance of environmental toxicants, targets of toxicant action
including the Ah receptor nonclassical receptors for endocrine
disrupting agents, and membrane bound heat shock proteins.

Membrane protein systems of interest to the National Institute of
Neurological Disorders and Stroke (NINDS) include receptors, ion
channels, structural proteins and other proteins involved in the
normal function and pathology of nerve cells in the central and
peripheral nervous system.  These include ion-selective channels
such as sodium, potassium, and calcium channels; ligand-gated ion
channels such as cholinergic, glutamatergic, gabaergic and
glycinergic receptors; compartmental and cytoskeletal elements
involved in protein trafficking and assembly, neurite growth, cell
adhesion, cell migration, synaptic structure, and vesicular
release; G protein-linked receptors; trophic factor receptors; and
transporters and pumps for ions, transmitters or macromolecules. 
Studies of conformational changes involved in normal function, in
the action of drugs, and in dysfunction of mutated proteins
associated with neurodevelopmental or neurodegenerative disorders
are of particular interest.  Protein-protein interactions important
for higher order structural assemblies and for pathological
associations characteristic of neurofilament, amyloid, Lewy body
and prion diseases are also of interest.

The above listings are not meant to be exclusive.  Structural
information obtained for any membrane protein will contribute to
understanding the general principles that underlie all membrane
protein structure and function.  Research on the non-membrane
embedded proteins associated with many of the cellular functions
listed above is also supported by the participating Institutes;
however, this program announcement is intended to emphasize the
need for additional research on structural aspects of the membrane-
embedded proteins involved in these processes.

APPLICATION PROCEDURES

Applications for R01 and P01 grants are to be submitted on the
grant application form PHS 398 (rev. 5/95) and will be accepted at
the standard application deadlines as indicated in the application
kit.  Application kits are available at most institutional offices
of sponsored research and may be obtained from the Division of
Extramural Outreach and Information Resources, 6701 Rockledge
Drive, MSC 7910, Bethesda, MD 20892-7910, telephone 301-710-0267,
Email [email protected].

Applications for the SBIR and STTR mechanisms are to be submitted
on the forms available in the Omnibus solicitations and will be
accepted on the SBIR and STTR application receipt dates.

Applicants planning to submit an investigator-initiated new (type
1), competing continuation (type 2), competing supplement, or any
amended/revised version of the preceding grant application types
requesting $500,000 or more in direct costs for any year are
advised that he or she must contact the Institute or Center (IC)
program staff before submitting the application, i.e, as plans for
the study are being developed. Furthermore, the application must
obtain agreement from the IC staff that the IC will accept the
application for consideration for award. Finally, the applicant
must identify, in a cover letter sent with the application, the
staff member and Institute or Center who agreed to accept
assignment of the application.  This policy requires an applicant
to obtain agreement for acceptance of both any such application and
any such subsequent amendment. Refer to the NIH Guide for Grants
and Contracts, March 20, 1998 at
http://www.nih.gov/grants/guide/notice-files/not98-030.html

The title and number of the program announcement must be typed in
Section 2a on the face page of the application (i.e., "Structural
Biology of Membrane Proteins," PA-99-004).

Submit a signed, typewritten original of the application, including
the Checklist, and five signed photocopies in one package to:

CENTER FOR SCIENTIFIC REVIEW
NATIONAL INSTITUTES OF HEALTH
6701 ROCKLEDGE DRIVE, ROOM 1040, MSC 7710
BETHESDA, MD  20892-7710
BETHESDA, MD  20817-7710 (for express/courier service)

REVIEW CONSIDERATIONS

Applications will be assigned on the basis of established PHS
referral guidelines.  Applications will be evaluated for scientific
and technical merit by appropriate scientific review groups
convened in accordance with the standard NIH peer review
procedures. As part of the initial merit review, all applications
will receive a written critique and undergo a process in which only
those applications deemed to have the highest scientific merit,
generally the top half of applications under review, will be
discussed, assigned a priority score, and receive a second level
review by the appropriate national advisory council or board.

Review criteria for STTR (R41 and R42) and SBIR (R43 and R44)
applications can be found in the omnibus solicitation references
above under Mechanism of Support.

Review Criteria for R01s and P01s:

The goals of NIH-supported research are to advance our
understanding of biological systems, improve the control of
disease, and enhance health.  In the written comments, reviewers
will be asked to discuss the following aspects of the application
in order to judge the likelihood that the proposed research will
have a substantial impact on the pursuit of these goals.  Each of
these criteria will be addressed and considered in assigning the
overall score, weighting them as appropriate for each application. 
Note that the application does not need to be strong in all
categories to be judged likely to have major scientific impact and
thus deserve a high priority score.  For example, an investigator
may propose to carry out important work that by its nature is not
innovative but is essential to move a field forward.

(1) Significance:  Does this study address an important problem? 
If the aims of the application are achieved, how will scientific
knowledge be advanced?  What will be the effect of these studies on
the concepts or methods that drive this field?

(2) Approach:  Are the conceptual framework, design, methods, and
analyses adequately developed, well-integrated, and appropriate to
the aims of the project?  Does the applicant acknowledge potential
problem areas and consider alternative tactics?

(3) Innovation:  Does the project employ novel concepts, approaches
or methods? Are the aims original and innovative?  Does the project
challenge existing paradigms or develop new methodologies or
technologies?

(4) Investigator:  Is the investigator appropriately trained and
well suited to carry out this work?  Is the work proposed
appropriate to the experience level of the principal investigator
and other researchers (if any)?

(5) Environment:  Does the scientific environment in which the work
will be done contribute to the probability of success?  Do the
proposed experiments take advantage of unique features of the
scientific environment or employ useful collaborative arrangements?
Is there evidence of institutional support?

In addition to the above criteria, in accordance with NIH policy,
all applications will also be reviewed with respect to the
following:

o  The adequacy of plans to include both genders, minorities and
their subgroups, and children as appropriate for the scientific
goals of the research.  Plans for the recruitment and retention of
subjects will also be evaluated.

o  The reasonableness of the proposed budget and duration in
relation to the proposed research.

o  The adequacy of the proposed protection for humans, animals or
the environment, to the extent they may be adversely affected by
the project proposed in the application.

AWARD CRITERIA

Applications will compete for available funds with all other
approved applications. The following will be considered in making
funding decisions:

o  quality of the proposed project as determined by peer review; o 
availability of funds;
o  program priority of research in the area of the program
announcement and other areas of Institute interest.

INQUIRIES

Inquiries are encouraged.  The opportunity to clarify any issues or
questions from potential applicants is welcome.

Direct inquiries regarding programmatic issues to:

Peter C. Preusch, Ph.D.
Division of Pharmacology, Physiology, and Biological Chemistry
National Institute of General Medical Sciences
45 Center Drive, MSC 6200
Bethesda, MD  20892-6200
Telephone:  (301) 594-5938
FAX:  (301) 480-2802
Email:  [email protected]

John C. Norvell, Ph.D.
Division of Cell Biology and Biophysics
National Institute of General Medical Sciences
45 Center Drive, MSC 6200
Bethesda, MD  20892-6200
Telephone:  (301) 594-0533
FAX:  (301) 480-2004
Email:  [email protected]

Richard W. Lymn, Ph.D.
Muscle Biology Program
National Institute of Arthritis and Musculoskeletal and Skin
Diseases 45 Center Drive, MSC 6500
Bethesda, MD  20892-6500
Telephone:  (301) 594-5128
FAX:  (301) 480-4543
Email:  [email protected]

Maren Laughlin, Ph.D.
Division of Diabetes, Endocrinology, and Metabolic Diseases
National Institute of Diabetes and Digestive and Kidney Diseases 45
Center Drive, MSC 6600
Bethesda, MD  20892-6600
Telephone:  (301) 594-8802
FAX:  (301) 480-3503
Email:  [email protected]

Jose Velazquez, Ph.D.
Chemical Exposures and Molecular Biology Branch
National Institute of Environmental Health Sciences
P.O. Box 12233, MD EC-21
Research Triangle Park, NC  27709
Telephone:  (919) 541-4998
FAX:  (919) 541-4937
Email:  [email protected]

Gabrielle Leblanc, Ph.D.
Division of Fundamental Neuroscience and Developmental Disorders
National Institute of Neurological Disorders and Stroke
Federal Building, Room 816, MSC-9170
Bethesda, MD  20892-9170
Telephone:  (301) 496-5745
FAX:  (301) 402-1501
Email:  [email protected]

Direct inquiries regarding fiscal matters to:

Ms. Phyllis Finch
Grants Administration Branch
National Institute of General Medical Sciences
45 Center Drive, MSC 6200
Bethesda, MD  20892-6200
Telephone:  (301) 594-5243
FAX:  (301) 480-1969
Email:  [email protected]

Ms. Nancy D. Curling
Grants Management Branch
National Institute of Arthritis and Musculoskeletal and Skin
Diseases 45 Center Drive, MSC 6500
Bethesda, MD  20892-6500
Telephone:  (301) 594-3503
FAX:  (301) 480-5450
Email:  [email protected]

Ms. Donna A. Huggins
Division of Extramural Activities
National Institute of Diabetes and Digestive and Kidney Diseases 45
Center Drive, MSC 6600
Bethesda, MD  20892-6600
Telephone:  (301) 594-8848
FAX:  (301) 480-3504
Email:  [email protected]

Mr. David Mineo
Division of Extramural Research and Training
National Institute of Environmental Health Sciences
P.O. Box 12233, MD EC-22
Research Triangle Park, NC  27709
Telephone:  (919) 541-1373
FAX:  (919) 541-2860
Email:  [email protected]

Ms. Tina Carlisle
Grants Management Branch
National Institute of Neurological Disorders and Stroke
7550 Wisconsin Avenue, Room 1004
Bethesda, MD  20892-9190
Telephone:  (301) 496-9231
FAX:  (301) 402-0219
Email:  [email protected]

AUTHORITY AND REGULATIONS

This program is described in the Catalog of Federal Domestic
Assistance Nos. 93.113, 93.821, 93.846, 93.847, 93.854, and.
93.859.  Awards are made under authorization of the Public Health
Service Act, Title IV, Part A (Public Law 78-410, as amended by
Public Law 99-158, 42 USC 285c-8 and 285k) and administered under
PHS grants policies and Federal Regulations 42 CFR 52 and 45 CFR
Part 74.  This program is not subject to the intergovernmental
review requirements of Executive Order 12372 or Health Systems
Agency review.

The PHS strongly encourages all grant and contract recipients to
provide a smoke-free workplace and promote the non-use of all
tobacco products.  In addition, Public Law 103-227, the Pro-
Children Act of 1994, prohibits smoking in certain facilities (or
in some cases, any portion of a facility) in which regular or
routine education, library, day care, health care or early
childhood development services are provided to children.  This is
consistent with the PHS mission to protect and advance the physical
and mental health of the American people.

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